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Chemical Senses Vol.12 no.l pp.

167-214, 1987

7th Congress of the European Chemoreception Research Organization


held in Davos, 2 2 - 2 6 September, 1986

Abstracts
/. Lectures The chemical senses: from microbes to man
Barry W.Ache C. V. Whitney Laboratory, University of Florida, St. Augustine, FL 32086, USA A broad, cross-phyletic consideration of the chemical senses reveals that, notwithstanding many speciesspecific differences, some rather striking similarities characterize chemoreception in diverse phyla. These similarities are not only confined to the molecular and cellular levels of organization, as might have been expected, but characterize more complex processes as well. Examples are presented which illustrate similarity in such diverse aspects of chemoreception as the multicomponent nature of chemical stimuli, the intermittent nature of signal detection, types of chemoreceptors, early events in the transduction process, principles of quality coding, central connections in the olfactory pathway, plasticity of response to odors and the association of chemoreception with immunorecognition. These species-independent features of the chemical senses may well reflect fundamental principles of chemosensory organization. Moreover, their existence gives further credence to the idea, first presented by Haldane (1954) and iterated since by others, that chemosensory competence in lower forms may have provided the evolutionary substrate upon which the chemical senses of higher organisms were formed. Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on February 16, 2014

Charm and other measures of human chemoreception


T.E.Acree
Cornell University, Geneva, NY 14456, USA Foods and other natural products comprise many different odor-active, simultaneously acting, stimuli to produce a single odor impression. Perception of odor is an integration of these separate responses into a single experience. Unfortunately, little is known about the nature of this integration, or, in the case of foods, which chemical components are involved. Attempts to understand the nature of odor using sensory analysis have not revealed the relationship between individual constituents and total perception. For example, descriptive analysis attempts to 'dis-integrate' perception into components by having people extract quality and intensity descriptions from their impressions of the whole. Charm analysis, on the other hand, is a technique that yields individual threshold measurements, free of inhibition or synergism. It can be used to determine the components of odor. In charm analysis, human subjects simply indicate the presence or absence of odor. The yes/no values produced by serial dilution of a sample are combined using c = D"-' where c, instantaneous charm, is equal to the dilution factor, D, raised to the power n 1, and n is the number of dilutions of the sample that produce an odor response. A charm response chromatogram is a plot of c versus time. These chromatograms can be produced without knowing what the odor-causing compounds are or how much are present in the food. Furthermore, the charm procedure yields measures of intensity without requiring panelists to make direct estimates of intensity. The panelists have only to decide whether or not they detect an odor. Therefore, panelists used in charm analysis can, with little training, produce accurate measures of relative odor intensity.

IRL Press Limited, Oxford, England

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ECRO VII: Abstracts

Discrimination by chemoreceptors (from fly to man)


V.G.Dethier Department of Zoology, University of Massachusetts, Amherst, MA 01003, USA Chemoreceptive systems must provide animals with means of discriminating stimuli quantitatively as well as qualitatively. Quantitative discrimination, measured by differential threshold (AT) is indicative of the smallest detectable change in stimulus intensity and represents the resolving power of the system. The curve describing AI in relation to the entire concentration range (I) is generally U-shaped; that is, discrimination is poor at low and high concentrations and best over the intermediate range. For chemoreceptors, from bacteria, fly, rat to man, the relationship has the same characteristics. This suggests the existence of a basic universal chemoreceptive mechanism. Experiments with bacteria and blowflies indicate that the mechanism derives from or is inherent in transduction at the receptor membrane. It is assumed that the relation between AI and I is dictated by the Law of Mass Action. Experiments with sugars show that for those that satisfy the Beidler equation discrimination is most acute in the concentration range around the dissociation constant. A change from a highly stimulating sugar to a weakly stimulating one shifts the AI/I curve to the right. Response biases also displace the curve. Among these are psychological biases (e.g. expectations, etc.) in man and metabolic variables in all animals. In flies and rats deprivation displaces the curve to the left. Finally, the basic curve is modified by integrating chemosensory information with information from other sources within the animals.

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Enzymes as models of receptors


Hans Dutler and Spartaco A.Bizzozero Laboratorium fUr Organische Chemie, ETH-Hdnggerberg, 8093 Zurich, Switzerland Enzymes and receptors share the property of binding a specific ligand. The two macromolecules differ in the way they 'convert' this primary process into function: enzymes catalyze a reaction whereby substrate is transformed into product and receptors undergo a change of structural properties and thereby alter the behavior of neighbour systems. Since both these processes are intimately related to the occurrence of specific interactions between the macromolecule and the ligand it seems reasonable to assume that the immediate response of the occurrence of these interactions is similar in nature for both and obeys the same laws. Thus knowledge of the molecular events taking place in the enzyme protein during substrate binding and reaction should provide relevant information as to how binding of a ligand to a receptor protein 'converts' a chemical signal into function. For example we have studied such processes in the case of peptides acting as substrates for proteases. The results of these studies allow us to track down in detail which amino acid residues in the enzyme protein are structurally affected due to substrate binding and reaction and how these molecular events are mediated from one residue to another. In another example we have been able to demonstrate such structural changes where a metal centre is involved.

Parallel neurophysiological and behavioral studies addressing the coding of gustatory quality information in mammals
Marion E.Frank Department of Oral Biology, University of Connecticut Health Center, Farmington, CT 06032, USA Early investigations into the neurophysiological basis for sensation in the gustatury system involved looking for representation of the taste qualities of sweet, salty, sour and bitter in electrophysiological recordings from neurons in the peripheral and central nervous systems. The qualities of taste sensations were identified psychophysically for humans; the nervous systems studied were mostly those of other mammals. Proposed neural codes were based on categorical neuron activation 0abclled line), level of interneuron activation (pattern) or distribution of neuron activation in time (temporal). More recent work has assessed the appropriateness of a theory of coding with comparisons of neural and behavioral measures from the same species. Anatomical, physiological and behavioral observations have identified striking species differences. The central taste pathways are distinct in different species; potent taste stimuli for one species are ineffective for taste neurons of other species; and taste stimuli of like quality to one species may have unlike qualities to another species. The assessment of species-specific taste quality with measurements of generalizations of conditioned flavor aversions has added a new dimension to behavioral analysis. Multivariate statistical analyses have helped identify the organization within large, complex neural and behavioral data matrices.

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ECRO VH: Abstracts Recent parallel neurophysiological - behavioral studies using these techniques have attempted to address taste quality coding at several levels of mammalian gustatory nervous systems. Adequate testing of coding theories awaits revaluation of the reliability and comprehensiveness of neural measurements and exact methods for quantitatively distinguishing among theories with precise predictions. Supported by a grant from the National Science Foundation (BNS85-19638).

Spatial and temporal aspects of molecular sensing


R.P.Futrelle
Northeastern University, Boston, MA 02115, USA Whether we deal with animals or simple cells, the same physical laws govern the motion of attractant molecules. Far from the organism, convection and turbulence move molecules. But close in, laminar flow and diffusion always dominate. Ultimately, the contacts the organism has with these molecules determine its directed motion. Using modeling and computer simulation we study some of the first stages of this transduction process. Our stochastic simulation models of the pheromone molecular motion near the insect sensillum [ Trends Neurosci., 7, 116-120 (1984)] are extended to look at transport to the sensillum interior. Similar techniques are used to model the relationship between receptor binding events and the establishment of directionality in Dictyostelium discoideum. We explain how the binding events can lead to the establishment of cell polarity via a reactiondiffusion model originally developed by Meinhardt and Gkrer. The results show a dose response for both the speed and accuracy of the cell's response, as well as fluctuations at very low attractant concentrations.

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Diet and the evolution of feeding strategies among primates in relation to chemoreception
C.M.Hladik
Musium-Ecologie, 4 Av-du Petit Chdteau, 91800 Brunoy, France Primate feeding strategies arc presented in the context of environmental conditions, including biochemical composition of potential foods. Actual measures (cf. several papers by D.Glaser et at.) and hypotheses concerning taste thresholds are related to the results of such field observations. Among Prosimians, although feeding behavior is the resulting combination of multiple factors, two dimensions at least have to be considered separately, (i) Specialization on peculiar food objects such as repugnant insects gums, nectars, insect secretions etc., in the genera Loris, Galago, Microcebus, Cheirogaleus, Euoticus and Phaner, involves specific taste tolerance and/or high sensibility to soluble substances, (li) Simultaneously, the maintenance of an efficient system of energy regulation requires different motivation patterns in genera such as Lemur, Lepilemur and Indri: accordingly a strategy of high energy input with high motivation can be opposed to a system of low energy expenditure using food items with a minimal energy content. Both strategies necessitate specific taste responses. Different simian species observed in natural conditions are subject to similar pressures of the biochemical environment. Examples will be given among genera Saguinus, Cebus, Macaca, Pan, Presbytis and Colobus, where behavioral flexibility complements a relatively rigid system of physiological adaptations. Human populations can also be examined under the particular angle of these environmental, physiological and behavioral adaptations. Recent field studies about taste thresholds in contrasted environments (tropical rain forest, arid zone and the arctic region) will illustrate these views and help to define the limits of biological impact in a cultural environment.

Information processing in the olfactory system: new findings to be integrated in our concept
Andre Holley University Claude-Bernard, 69622 Vdleurbanne Cedex, France Our concept of information processing and odour discrimination mechanisms in the first stages of the olfactory system must be periodically re-examined in the light of new available evidence. This presentation attempts to do this, focusing on experimental findings obtained in our laboratory following two different

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ECRO VII: Abstracts approaches, (i) Studies by Bouvet and Delaleu showing that anti-dromic stimulation of the ophthalmic branch of the trigeminal nerve, like neuroepitbelial superfusion with substance P, alter the spontaneous activity of receptor cells distributed in relatively large areas of the neuroepithelium. (ii) Systematic investigations by L.Astic and D.Saucier of the receptor cell projections to the olfactory bulb, which demonstrate that small groups of glomeruli are innervated by receptor cells distributed in relatively large areas of the neuroepithelium.

Behavioral studies in humans and animals on 'umami'


Y.Kawamura Furuc-dai 3-24-12, Suita City, Osaka 565, Japan Biochemical, food science, neurophysical, psychophysical and behavioral studies in humans and animals on umami, that is, the taste of glutamates and 5'-nucleotides, are reviewed. Taste receptors of amino acids are proteins, and in a number of systems, the receptor for L-glutamate (Glu) is unique and not shared by other amino acids. In rats, the presence of 5'-nucleotides enhances the affinity of the taste receptor for Glu; in bovine circumvallate papillae, it increases the maximal capacity of Glu binding. Monosodium glutamate (MSG)-responsive single-unit fibers are generally unresponsive or only slightly responsive to the basic taste stimuli represented by NaCl, HC1, sucrose and quinine. Multidimensional scaling of taste evaluations by Japanese and American subjects suggest that umami lies outside the tetrahedron spanned by four basic tastes. Umami makes an important contribution to the natural flavor of crab meat, and Glu content of meats increases with aging. Hawaian infants show a preference for umami taste. Flavor preferences in both rats and humans are affected by protein status, and data suggest that umami could be a marker for adequate protein status.

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Chiral flavor substances - analysis and structure-activity relationships


A.Mosandl Lehrstuhl fUr Lcbensmittelchemie, Universit/U Wdrzburg, Am Hubland, D-8700 WQrzburg, FRG After a short introduction to the problems of sample preparation procedures as an important step of flavor analysis, the principle methods of chiral flavor analysis are presented. By means of selected examples, both of analysis using achiral phases after derivatization and of direct analysis with chiral phases, actual techniques are demonstrated. The advantages and disadvantages of the different techniques as well as structureactivity relationships are discussed using recent results from our group.

Pleasure and disgust: some intercultural evaluations of odour experiences


M.Schleidt Fondumgsstclle fir Humanethologie, Max-Planck-lnstitut fir VerhaUensphysiologie, D-8131 Seewiesen, FRG In evaluating odours, subjective conditions play a large role. Odour experiences are less oriented towards the gaining of cognitive knowledge than is the case in experiencing sight and sound. Furthermore, they stimulate emotions to a presumably much higher degree than the two other sensory qualities. Because emotions provide sustaining motives for future actions, odour experiences are important in the effective evaluation of situations. Odours can characterize special situations like signals. For this task they are well equipped not only through their close affective connections, but also through their long lasting storage in long-term memory. Intercultural comparison, which was done mainly on the basis of odour memories of informants, showed that the relationship of subjective conditions to odour qualities is not quite optionally variable per individual, but relatively uniform throughout the groups. The subjects evaluated most of the odours, which they named out of all realms of their life and the total environment, either as positive or as negative. In this evaluation of odours the meaning of the whole situations, signalled by these odours, is important and indicates harmful or beneficial conditions. Odour works as a signal device for the satisfaction of basic human needs in the physical, mental and social domain. The analysis revealed that odour-released emotions trigger specific actions, mainly to or away from the appertaining situations. In general odour stimuli seem especially well suited to regulate human actions via effect.

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//. Seminars 1. Pollution and Chemoreception Research Sensory responses to indoor air pollution
William S.Cain John B. Pierce Foundation Laboratory and Yale University, New Haven, CT 06519, USA Over the last half-century, engineers have recognized that ventilation requirements in buildings need to derive from psychophysical judgments of the quality of the indoor air. Recent research in this laboratory has focused on two 'perceptual contaminants', body odor and tobacco smoke odor. These form two ends of a continuum of severity; the odor generated by tobacco smoke greatly exceeds that generated by simple occupancy. Because of its high odor and because tobacco smoke odor is a more stable contaminant than body odor, smoking occupancy requires much greater ventilation than non-smoking occupancy. The ventilation rates necessary to achieve comfort are almost too high to be achieved, even when the judges comprise a mixed group of smokers and non-smokers. When the judges comprise only non-smokers, ventilation rates for comfort during smoking occupancy are in fact impossible to achieve. This situation indicates a pressing need for mitigation strategies. Filtration of particles from the cigarette aerosol offers one important possibility. Unfortunately, electrostatic precipitation of the particles had essentially no effect on the odor experienced by visitors to a smoking environment and had only a small effect on the odor and irritation experienced by the occupants in the space. In other laboratories, the use of Cambridge filter pads to trap the aerosol has mitigated irritation substantially. This outcome suggests that some vapor phase constituent(s) trapped by Cambridge filters and unimpeded by electrostatic precipitation may hold an important key to mitigation.

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Standardisation on olfactometry with respect to odor pollution control


M.Hangartner Institute of Hygiene and Applied Ergonomics, ETH Zurich, Switzerland In many European countries clean air acts were introduced in the last two decades and consequently the interest in regulating odor pollution has increased. For regulatory purposes measuring methods must be provided, which give comparable results and are accepted both by administration and odor-producing industry. The method most commonly used to control odorous emissions is odor threshold determination, expressed in dilution to threshold numbers. Based on different national guidelines (FRG, F, N, UK) recommendations on olfactory measurements were worked out during a EEC/FAO workshop in Silsoe (UK) in 1985. The main points concern sampling of odorous air, olfactometers, panel, operational procedure and presenting of results. Under the supervision of VDI (Association of German Engineers) several inter-laboratory tests were conducted to check the effect of the VDI guideline 3881 'Olfactometric method of odour threshold determination-fundamentals' on the variation and transferability of results by means of defined odorants. The ratios between the highest and the lowest measuring value per substance were 510 and the ratios between the upper and lower limit of the 95 % confidence interval 2 3. These are considerably inferior to those found in earlier olfactometric inter-laboratory tests. The result shows no homogeneous odorant-independent sequence among the participating laboratories. Likewise for other sensory properties, such as intensity and hedonics, guidelines will be prepared. Most of all, thefindingsof research should be accessible to the engineer, even if there are some compromises involved. The purpose of this introduction is to stimulate the discussion of existing methods for common use and the recommendation of other methods, which should lead to further guidelines that can be fruitful to the public.

Odor intensity measured by an audio method


Bonnie King-van der Pers Naarden International, PO Box 2, 1400 CA Bussum, The Netherlands A non-verbal, non-numerical method for measuring odor or taste intensity by means of cross-modal matching with pitch is described. The device with which the measurements are obtained is called an intensometer.

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ECRO VII: Abstracts and it allows subjects to simply react to intensity without having to describe or quantify what is perceived. The use of this device is illustrated by presenting data for odor intensity functions from flavor compounds. Individual as well as panel data are discussed. The method is compared wtih those traditionally used to obtain odor intensity functions.

Standardized olfactory tests in Japan - a review over ten years


Sadayuki F.Takagi Department of Physiology, School of Medicine, Gunma University, Maebashi 371, Japan A research group of 14 members from 13 universities was organized in 1971, aiming to manufacture a standardized olfactometer. At first, 10 odorants were selected as standard test odors. By repeating 10-fold dilution, a series of test solutions from 10~' to 1 0 " " was made for each of the 10 odors. Detection and recognition thresholds for these solutions were sought among many healthy men and women 18-25 years old at the 13 universities. After analysis of the test data, five odors were selected among the 10. They were: /3-phenyl ethyl alcohol (A), methyl cyclopentenolone (B), iso-valeric acid ( Q , 7-undecalactone (D) and Skatole (E). Averages of the detection thresholds to these odors were calculated. Repeating 10-fold multiplication or 10-fold division of these averages, a new series of odor solutions from 10~2 to 105 was made for the five odors. The solutions were bottled and set in a metal frame. This newly manufactured 'T & T olfactometer' was endorsed in 1975 by the Otorhinolaryngology Society of Japan. The olfactogram was prepared for recording the results of clinical tests with this olfactometer. A venous olfactory test was also standardized, using Alinamine (thiamine propyl disulfide) as a test odorant. Since then these tests have been widely used in Japan. In 1983, the Ministry of Welfare in Japan approved a test with the T & T olfactometer and assigned 420 social welfare reimbursement points (at 12 Yen per point) for it.

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Role of olfactometry and annoyance assessment in dealing with odor pollution


G.Winneke and J.Kastka Medical Institute of Environmental Hygiene, University of Duesseldorf, D-4000 Duesseldorf, FRG The perception of odors in the environment is frequently associated with negative emotional responses. The regulation of environmental odor pollution is hampered by the difficulties of analytical assessment of complex chemical mixtures at low atmospheric concentrations, by the sensitivity of the sense of smell, as well as by the subjective nature of the emotional response to olfactory stimulation. Olfactometry and annoyance assessment by means of population surveys have, thus, been used for the monitoring of environmental odor pollution, for the setting and control of standards for odorous compounds, for planning purposes, or for the assessment of the efficiency of abatement programs. The organoleptic determination of odor concentrations in terms of dilution ratios is most frequently used to characterize emissions at the stack, and to predict odor concentrations in the neighbourhood by means of dispersion models. Atmospheric odor concentratons can, within certain limits, be measured as well. Typical concentrations measured near various industrial sources by means of a mobile olfactometric laboratory are given. A rational approach for the setting of permissible standards for environmenta] odor pollution would require satisfactory correlations between environmental odor concentrations and annoyance responses from the exposed population. Such correlations, if significant at all, are typically low. One reason is the interference of intensity and hedonic tone of different odor emissions. Examples are given showing that odor annoyance near different industrial sources, as assessed by questionnaire techniques, varies markedly despite comparable atmospheric odor concentration profiles. The implications of these findings for the regulation of environmental odor pollution are discussed.

2. Progress and Knowledge of Receptors Taste receptor site specificity for amino acids
Joseph G.Brand1-21, D.Lynn Kalinoski1, Bruce P.Bryant1 and Richard Bruch1 l Monell Chemical Senses Center, 3500 Market Street, and *Veterans Administration Medical Center and 'University of Pennsylvania, Philadelphia, PA 19104, USA For the past decade, our laboratory has used the cutaneous taste system of the catfish, Ictalurus punctatus, as a model for studying peripheral biochemical steps in taste transduction. We have taken advantage of the

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ECRO VII: Abstracts specificity shown by this animal for amino acid taste stimuli in characterizing the initial receptor event in taste. We have also exploited monoclonal antibody technology to generate competitive agonists/antagonists to the receptor event. Both biochemical and electrophysiological techniques have been used to investigate: (i) the extent to which the enantiomeric stimuli, L- and D-alanine, interact with both unique and common receptor/transduction processes and (ii) the specificity of the receptors) for the potent stimulus, L-arginine. The J ^ p p for binding to Fraction P2 from taste epithelium was 1.5 ^M for L-alanine and 25 jiM for Dalanine. The enantiomers competed with one another for binding, although the lower affinity sites for Dalanine were less susceptible to L-alanine inhibition than was the high affinity site for D-alanine. Recordings from most nerve bundles indicated that L- and D-alanine cross-adapted in an electrophysiological assay although this cross-adaptation was not always complete. L-alanine was the more potent stimulus of the enantiomeric pair. L-Arginine was bound to a high affinity site with a Kr*pp of 0.37 pM and to a lower affinity site with a Afpjpp of 13 pM. Binding to only the lower affinity site could be inhibited by L-lysine, D-arginine and L-a-amino-j3-guanidino-propionic acid (L-AGPA), but no inhibition of either site was observed using Lglutamate, glycine or L-alanine until high levels were achieved ( > 10~3 M). Electrophysiological studies demonstrated that only L-arginine and L-AGPA were effective stimuli. Only L-AGPA was an effective crossadapting stimulus to L-arginine. The high affinity site for L-arginine thus appears very specific. The monoclonal antibody, G10, developed in our laboratory (Goldstein and Cagan, Proc. Natl. Acad. Sci. USA, 79, 7595), is a competitive inhibitor of L-alanine binding. Using G10, immunoblots of taste plasma membrane proteins separated by SDS-PAGE revealed a major stained band, absent in other tissues tested. This band at 110 000 daltons, was positive to concanavalin A and wheat germ aggluunin. These studies will lead to molecular characterization of separate taste receptor classes. Their specificity and ability to react with monoclonal antibodies can be exploited in isolation and purification of receptor macromolecules. Supported in part by NIH Research Grants NS-15740 and NS-22620, by NRSA Fellowship F32 NS-07809 and by the Veterans Administration.

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Role of GTP-binding regulatory proteins in receptor-mediated phosphoinositide turnover in olfactory cilia


Richard C.Bruch, D. Lynn Kalinoski and Taufiqul Huque Monell Chemical Senses Center, 3500 Market Street, Philadelphia, PA 19104, USA GTP-binding proteins (G-proteins) were identified in isolated olfactory cilia from the channel catfish (1ctaluruspunctatiLS) by immunoblotting and bacterial toxin catalyzed ADP-ribosylation. The common 0-subunit of the G-proteins was identified by immunoblotting. A cholera toxin substrate, mol. wt 45 000, which may correspond to the stimulatory G-protein G,, was also identified in the isolated olfactory cilia. In addition, the cilia contained a single pertussis toxin substrate, mol. wt 40 000 which also cross-reacted with antiserum to the common sequence of G-protein a-subunits [Mumby el al. (1986) Proc. Nail. Acad. Sci. USA, 83, 265]. The role of G-proteins in mediating odorant-sensitive phosphoinositide turnover [Huque and Bruch (1986) Biochem. Biophys. Res. Commun., 137, 36] was investigated by examining the effects of exogenous guanine nucleotidcs on receptor binding activity and on phospholipase C (EC 3.1.4.11) activity. The L-amino acids, alanine and arginine, are effective olfactory stimuli for the catfish in electrophysiological assays which do not cross-adapt significantly [Caprio and Byrd (1984) /. Gen. Physioi, 84, 403]. These amino acids bind specifically to separate receptors in the isolated olfactory cilia with similar affinity (Kd 10~* M). Addition of GTP or its non-hydrolyzable analogue Gpp(NH)p to the binding assay mixture decreased the affinity of both receptors by about one order of magnitude, indicating that these receptors are coupled to a G-protein. Phospholipase C activity was stimulated to varying extents in the presence of odorant amino acids. The stimulation was dependent on odorant concentration and time of exposure. The enzyme activity was also stimulated in a similar manner by guanine nucleotides. A synergistic, nearly additive, stimulation of the enzyme activity was observed in the simultaneous presence of odorants and GTP. These results indicate for the first time that olfactory receptors are coupled to a G-protein which mediates phosphoinositide turnover. These results also suggest a possible role for this pathway in olfactory transduction. This work was supported in part by BRSG SO7-RRO5825-O7 from the Biomedical Research Support Grant Program, National Institutes of Health and in part by a grant from the Veterans Administration to J.G.Brand.

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Stimulus-evoked transmucosal currents in taste and olfactory reception


John A.DeSimone Department of Physiology and Biophysics, Virginia Commonwealth University, Richmond, VA 23298, USA Two of the special senses, taste and olfaction, are devoted to the detection of exogenous chemical substances. In each case the initial process of stimulus recognition is believed to occur in the apical membranes of the receptive sensory elements. The set of molecular events which constitute stimulus recognition and subsequent sensory cell transduction is extremely complex, but recent biophysical and biochemical studies have yielded new insights regarding transduction in both senses. In the case of mammalian NaCl taste, experiments give evidence of two separate receptor mechanisms. One of these is an amiloride blockable sodium channel. In this case the molecular receptor is an ion specific channel and depolarization of the receptor cell probably results from the direct transport of sodium ions across the apical cell membrane. Repolarization would then occur through activation of the sodium pump. By fitting a single-sided Ussing chamber to the anterior rat tongue, we have recorded changes in either the open-circuit potential or the short-circuit current under NaCl stimulation while monitoring the integrated chorda tympani response. Under short-circuit, stimulation with NaCl results in a rapid inward current which coincides with increasing neural activity. This is followed by a slow inward current which coincides with neural adaptation. The early rapid inward current is reduced by amiloride as is the neural response. The slow current component relaxes exponentially with the same time constant as that of neural adaptation. The results suggests that receptor cells are depolarized by sodium entry and repolarized by increased sodium pump activity. In the case of olfaction we have measured odorantevoked current transients across the frog olfactory epithelium under short-circuit in vitro. The response consists of a rapid inward current followed by a slower return to baseline when the odorant is delivered as a bolus. The response is increased by mkromolar concentrations of 8-Br-cAMP, forskolin, and phosphodiesterase inhibitors. Under the same conditions 8-Br-cGMP is a strong inhibitor of the response. The results indicate that the current transient represents the odorant-gated transduction current and is regulated via a G-protein activated adenylate cyclase as recently proposed by Pace et al.. Ion replacement studies show that the inward current transient is mainly due to sodium. Supported by NIH-NS13767.

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New ligands for the pyrazine binding protein


P.Pelosi and R.Tirindelli Istituto di Industrie Agrarie, via S.Michele, 4, Pisa, Italy The specificity of the pyrazine binding protein from cow nasal tissue has been measured with 50 compounds, using competitive binding experiments. The method involved incubation of the purified protein with labelled 2-isobutyl-3-methoxypyrazine and each ligand at three different concentrations, gel chromatography on G-25 microcolumns and evaluation of the bound fraction. The compounds tested were: (i) a series of odorants of ~ 10 carbon atoms, bearing some structural relation to known good ligands: they included cyclic ketones and alcohols, such as methanol, carvone, camphor; (ii) compounds known as strong odorants, but of different chemical structure, such as geosmin, straight chain aldehydes, androstenone, and others; (iii) molecules containing two pyrazine rings linked by a saturated chain of 48 carbon atoms; (iv) compounds obtained by connecting two molecules of 4-butyl-5-propyl-2-amino thiazole with chains of various length; (v) fluorescent derivatives of 4-butyl-5-propyl thiazole; (vi) affinity labels, bearing bromoester or azide groups, both on pyrazine and thiazole rings, and synthesized for the first time. The results have shown that several of the compounds tested bind to the protein as well as the labelled pyrazine and some of them with even higher affinity. Some of the fluorescent compounds and the affinity label derivatives are strong ligands and can be used for specific studies on the protein.

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///. Special Session on Chemoreception in Primates 1. Taste Visualization of binding sites for the sweet-tasting protein, thaumatin, in taste pores of monkeys and rats
A.I.Farbman, C.K.Ogden-Ogle, G.Hellekant1, R.M.Albrecht1 and H.van der Wei1 Department of'Neurobiology and Physiology, Northwestern University, Evanston, IL 60201, and ^Department of Veterinary Science, University of Wisconsin, Madison, WI 53706, USA Taste pores in monkeys and rats contain the apical processes of epithelial cells constituting the taste bud. Some of these processes are narrow microvilli, others are broad, club-shaped extensions. We conjugated the sweet-tasting protein, thaumatin, to 5 or 10 nm colloidal gold particles. Rhesus monkeys and rats were treated with atropine to inhibit salivary secretion, and then anesthetized. After a thorough rinse of the tongue surface, the gold-labeled protein solutions were applied in either the foliate or vallate papilla regions for ~ 3 min. Biopsies of the taste bud-containing papillae were fixed and processed for electron microscopy. The gold particles are easily visualized with the electron microscope and served as a marker for the presence of the protein. As controls, we used albumin and acetylated thaumatin, both labeled with gold; neither of these tastes sweet. Electron microscopic examination of taste bud secretions from both species revealed many small (50-100 nm) extracellular membrane bounded vesicles of cytoplasm that appear to have been produced by an active apocrine secretory process or blebbing from the microvilli. There is evidence, moreover, that parts of the broad clubshaped processes extending into the taste pore are pinched off and these fragments of cytoplasm arc extruded into the oral cavity. We found gold particles in the pores, associated primarily with 50-100 nm vesicles. In a few instances, we saw gold particles concentrated on a small (~80 nm) bleb-like protrusion on the surface of a microvillus. There were very few particles associated with the larger apical processes or their detached fragments. We found few gold particles bound to the surface of the tongue in non-gustatory regions, i.e. distant from taste pores. Control specimens treated with gold-labeled albumin or acetylated thaumatin showed very few panicles in the pore. We conclude that the binding of thaumatin to certain stuctures in the taste pore has a relatively higher affinity than binding elsewhere on the tongue surface. The specificity of the binding to the 50-100 nm membranous vesicles suggests the possibility that the ligand is bound to receptor regions on these membranes, which have been discarded from taste microvilli. Supported by USPHS grant # NS-17021.

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Effect of gymnemic acid on the sweet taste in apes


D.Glaser, H.van der Wei1 and G.Hellekant2 Anthropologisches Institut, UniversitatZurich, Switzerland, lvan Beethovenlaan 5, NL-3161 RVRhoon, The Netherlands, and department of Veterinary Science, University of Wisconsin, Madison, WI 53706, USA Gymnemic acid (GA) suppresses the sweet taste in man. In three non-hominoid primate species this effect was not noticed by electrophysiological studies. The hominoid species chimpanzee, however, showed in their electrophysiological response a decrease of 75% to sucrose as well as a complete abolition to acesulfam-K, aspartame, monellin and thaumatin. The first preliminary behavioural experiments with six hominoid species indicated no GA effect on the preference for sucrose. In the present study, the effect of GA on the sweet taste sensitivity was investigated in more detail. Two-bottle preference tests, using different concentrations of sucrose, thaumatin and aspartame were carried out. All the six hominoid species demonstrated a clear preference for sucrose, thaumatin and aspartame solutions to water. In the siamang (Symphalangus syndactylus) no effect of GA was observed. In the gibbon (Hylobates pileatus), another small ape, there was an effect on the sucrose intake but not on thaumatin and aspartame. In the chimpanzee (Pan troglodytes troglodytes) there was a slight depression of sucrose consumption and one animal rejected aspartame completely. In the gorilla (Gorilla gorilla gorilla) some suppression of the intake of the three sweeteners was observed after GA. The difference of the GA effect in the two orang-utan species was remarkable: Pongo pygmaeuspygmaeus showed practically no effect after GA, in contast to Pongo pygmaeus abelii where thaumatin and aspartame were completely suppressed by GA.

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Neural recordings of taste modifiers and sweet taste in primates


G.HeUekant, J.N.Brouwer, A.I.Farbman, D.Glaser, C.HSrd af Segerstad, T.W. Roberts and H.van der Wei University of Wisconsin. Madison, Wl 53706, USA Some substances, which have little taste or smell of their own at the concentrations used, affect the taste quality of other compounds. These can be called taste modifiers. Miraculin and gymnemic acid are powerful examples. An overview of the effects of miraculin is given. The emphasis is on the neurophysiology but also includes related behavioral and psychophysical data and some discussion of its hypothetical mode of action. Gymnemic acid represents another aspect of taste modifiers with its sweet depressing or blocking effects. Here again differences in primate neurophysiological reactions have been observed and are presented. The remarkable phylogenetic relationship of these two compounds is included. The sweet taste of monellin and thaumatin, as it is mirrored in neurophysiological recordings of certain primates, is reviewed. Some ideas of their mode of action and structure are discussed. The phylogenetic relationship of monellin and thaumatin is known, but there are other potent sweetners like aspartame which show similar features. Some data from recordings of the response to other sweeteners are presented and related to observations in humans.

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Chorda tympani nerve responses to intra-arterial and surface stimulation of the tongue in rhesus monkey and rat
G.HeUekant, H.van der Wei, J.N.Brouwer, D.Glaser1, C.Hard af Segerstad and T.W.Roberts Department of Veterinary Science and Wisconsin Regional Primate Center, University of Wisconsin, Madison, Wl 53706, USA, and 1Anthropologisches Institut, UniversitOt Zurich, Switzerland The effects of intra-arterial injection of monellin, thaumatin and miraculin on the activity of the chorda tympani proper nerve have been recorded in the monkey and rat. The substances were injected into the blood stream at the branching of the lingual artery. It was found that monellin and thaumatin elicited a nerve response in the monkey but not in the rat. Miraculin had no effect in either species. It is concluded that the response to intra-arterial injection in the monkey was caused by stimulation of its taste buds by monellin and thaumatin. The results suggest that there are receptors to these two compounds also on the parts of the taste cells not facing the oral cavity.

Taste processing in the nucleus of the solitary tract and in three cortical taste areas in primates
Edmund T.Rolls Oxford University, Department of Experimental Psychology, Oxford, UK First, analysis of the activity of 360 single neurons in the gustatory pathways in primates (cynomolgus monkeys) showed that the tuning of neurons becomes sharper as information progresses through the taste system. For example, the breadth of tuning (1) to the four prototypical stimuli 1.0 M glucose, 1.0 M NaCl, 0.001 M quinine HC1 and 0.01 M HC1, and 20% blackcurrant juice, of neurons in the first central relay, the nucleus of the solitary tract was 0.87 0.01 (mean sd), 0.67 0.02 in the frontal opcrcular (primary) taste cortex, 0.56 0.03 in the insular taste cortex and 0.39 0.23 in a new caudolateral orbitofrontal cortex (secondary) taste area. Second, it was found that feeding the monkey to satiety with glucose had no effect on gustatory responses of neurons in the nucleus of the solitary tract, the frontal opercular taste cortex or the insular taste cortex, but decreased the magnitude of the neuronal responses of orbitofrontal cortex taste neurons which responded to glucose to zero. Third, the responses of orbitofrontal cortex taste neurons were shown to decrease to foods on which the monkey was fed to satiety, but to continue to foods which had not just been eaten, that is they reflected sensory-specific taste satiety. It is proposed that tuning becomes sharper in the taste sytem through unimodal processing stages so that after this processing satiety can operate with some specificity, allowing responses to foods eaten to decrease without producing a decrease in responsiveness to other gustatory stimuli, and so that associations can be made to other modalities with minimal interference in an association memory (2). (1) Smith.D.V. and Travers.J.B. (1979) Own. Senses, 4, 215-229. (2) Rolls.E.T. (1986) InChangeux,J.-P. and Konishi.M. (eds), Neural and Molecular Mechanisms of Learning. Springer-Verlag, Berlin.

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2. Olfaction Olfaction and social communication in callitrichid monkeys


G.Epple1-2, A.Belcher " , I.Kuderling1 and A.B.Smith.IlP ] ^German Primate Center, 34 Gottingen, FRG, 1Monell Chemical Senses Center, 3500 Market St., Philadelphia, PA 19104, and ^Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA Scent marking with specialized skin glands and urine is common in all callitrichid monkeys. Behavioral studies on several species have suggested that in this primate family, scent marking and the chemical signals contained in the marks are important in a number of sexual and social situations. Our comparative studies on saddle-back and cotton-top tamarins have shown that the scent marks of both species contain a number of communicatory messages. In both species the scent marks are chemically complex, consisting mainly of secretions from apocrine and sebaceous skin glands and of urine. However, other ingredients, such as compounds derived from the genital tract, may also be important. In saddle-back tamarins, the major volatile constituents of glandular secretions and of the complex natural scent markers from males and females are squalene and 15 esters of butyric acid, but low concentrations of numerous other volatiles as well as compounds of higher mol. wt are also present. Studies with two subspecies have shown that concentration patterns of some of the butyrates are diagnostic of subspecies, while concentration patterns of other butyrates are diagnostic of gender, regardless of subspecies. Bioassays of fractions and synthetic mixtures have suggested that the butyrates are important but not sufficient for encoding subspecies specificity in the scent marks, and that yet unidentified synergists are also necessary. In cotton-top tamarins, the major volatile constituents of natural scent marks are squalene and cholesterol. In addition, a number of unidentified volatiles are present in low concentrations. Secretion manually expressed from the subrapubic part of the glandular pad contains all 15 butyrates previously identified in saddle-back tamarins, but natural scent marks contain only trace amounts of these esters. Differences in the manner in which both species use the suprapubic and circumgenital portions of their glandular pads during scent marking may account for this finding. The results of our behavioral and chemical studies show that the butyrates are secretory products of the skin glands of both species and suggest that they may play a differential role in encoding communicatory signals. Supported by grants from the National Science Foundation (BNS 83-00937), the National Institutes of Health (RO1 NS21790) and the Deutsche Forschungsgemeinschaft.

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Development of chemo-communication mechanisms among non-human primates


C.S.Evans
Department of Biological Sciences, Glasgow College of Technology, Glasgow, UK Analysis of the differential importance of early social experience previously stressed non-olfactory cues as determinants of later behaviour. Infants of Prosimians and New-World monkeys combine early scent-marking with the discrimination of maternal odours. The primary olfactory system persists, but varies in its importance; while the secondary component of the mammalian dual system is only selectively retained within the group. Prolonged parental contact even in (relatively) precocial primates allows a substantial chemoreceptive contribution to behavioural adjustment. The scent cues from a young primate could provide (or omit) 'identifying' chemo signals, for use by related/unrelated adults. Assessment of early olfactory-led learning is handicapped by incomplete developmental timetables: often structural and behavioral ontogeny remain uncorrelated. Early plasticity of odour response may, as in rodents, relate to structural changes in both pathways. A survey of the status of the vomero-nasal organs among 'lower' primates indicates considerable variation in the maturational speed of post-natal neurogenesis. Neonatal functional dependence on VN olfaction is low or absent in some Lorisidae, Lemuridae and Callithricidae. If extensible to other families this provides a model for early provision of chemosensory input from either, or both components of the dual system. In the pre-pubertal period reproduction and its adaptive mechanisms are a likely target for dependence on such input. Comparison with rodents readily suggests several maturational roles for olfaction, particularly as a determinant of adult social/endocrine status.

Occurrence of sternal and axillary glands in hominoids


T.Geissmann Anthropological Institute, University Zurich-Irchel, CH-8057 Zurich, Switzerland Although glandular structures in the sternal region are known to occur in many primates species, in hominoids such a gland is only known of in the orang-utan. In the latter, the gland has been thought to be in a stage of

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ECRO VD: Abstracts regressive evolution. This report presents evidence for the occurrence of a sternal gland in at least two species of gibbons (Hylobates syndactylus and H. agilis). In contrast to the sternal gland, the axillary glandular organ is present only in the hominoid genera Gorilla, Pan and Homo, none of which is reported to possess a sternal gland. Attention is drawn to the complementary occurrence of sternal and axillary glands in hominoids. It has repeatedly been proposed that the sternal gland in primates might be a retained primitive feature, whereas the axillary gland of man and the African apes has been interpreted as a derived, synapomorph characteristic. The possibility is discussed here that disappearance of the sternal gland and appearance of the axillary gland occurred not only in temporal, but also in functional, interconnection during hominoid evolution.

Chemical signals and the reproductive capacity of a male Prosimian primate


A.Schilling and M.Perret Laboratoire d'Ecologie, 4 av. du Petit Chateau, 91.800 Brunoy, France Some Prosimian primates, especially nocturnal forms, like Cheirogaleidae or Lorisidae, have retained chemoreceptor systems as well developed as in Insectivora. These systems allow a keen discrimination ability but it is not known if they are involved in the sensory mediation of behavioral or physiological reproducible effects as has been shown for rodent reproduction. In order to investigate the following questions, we have focused this research on male Microcebus murinus (Cbeirogaleidae), which display both a clear seasonal cycle of activity and a clear hierarchical social structure when kept in heterosexual groups. (i) Are behaviors involved with chemical communication such as marking behavior related to the seasonal increase of social activity in the mouse lemur? (ii) Do the scent marks, produced during this marking activity, have any effects on subordinate conspecifics? If so, can we assume that the factor responsible for these effects is contained in die excreted urine of the emitter and that it acts upon the receiver under a volatile form? (in) What do we presently know about the characters of this chemical factor: (a) Absence in non dominant subject urine, (b) Pheromonal-like type of action, (c) Presence in the organic phase alone of the urine, (iv) Which endocrinological mechanisms, activated by the central neural processing of this factor, may account for the inter-male sexual inhibition? (a) Glucocorticoids. (b)Prolactin. Although many questions concerning the emission or the reception of the mouse lemur urinary signal remain to be clarified, these results illustrate that in some Primates chemical communication is effective at an infracortical level. If this physiological stress effect occurs also in naturally behaving male Microcebus murinus, then several characteristics of the reproductive and social behavior of this primitive primate species would be better understood. Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on February 16, 2014

Chemical signals in the context with aversive behavior in tupaias and primates
F.v.Stralendorff Department of Animal Physiology, University of Bayreuth, UniversitOtsstr. 30, D-8580 Bayreuth, FRG Chemical signals may be connected with aversive behavior in different ways: firstly as chemical signals which cause aversive behavior and secondly as metabolic products released by animals responding aversively to a particular stimulus. On the basis of bibliographic investigations and on my own observations I review circumstantial evidence for the occurrence of such signals. Within this scope aversive behavior is analyzed, the biogenesis of presumed signal substances is discussed with regard to its control mechanism and considerations are made on how the signal receiver can 'understand' the information transmitted. Lastly the question is raised: What is the adaptive significance of those signals and to what degree do they play a role in the communication of higher primates including men?

IV. Workshop on Chemoreception in Human Behaviour Chemoreception and feeding behavior, appetite, anorexy
France Bellisle Laboratoire de Newobiologie de la Nutrition, Collige de France, Paris, France Chemoreception is an essential part of the complex learning mechanism through which food preferences develop, and through which eating behavior is determined. The development of individual food preferences

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ECRO VII: Abstracts has been extensively studied but remains poorly understood. The impact of stated or measured preferences on actual consumption is also being studied experimentally. Both palatability as such and variety of flavors exert predictable effects on intake and on the micro-structure of meals. An increase in palatability leads to accelerated ingestion patterns; during ad libitum mixed meals, human subjects' choices tend to optimize sensory stimulation. Much research has been devoted to comparisons between normal weight, healthy humans versus obese or anorectic persons. Although it is impossible to generalize any one behavioral observation to whole populations, recent experiments suggest certain mechanisms that could come into play in certain hyperphagic or hypophagic persons with intact chemoreception. Hyperresponsivity to food stimulation is suggested by an analysis of the micro-structure of meals in certain obese persons. Chronically food-deprived individuals ('restrained eaters') also present signs of disturbed food intake control. In anorectics, denial of the positive valence of foods is such that perception of the sensory characteristics of food stimuli can be affected.

Chemoreception in human behaviour: experimental analysis of the social effects of fragrances


M.D.Kirk-Smith and D.A.Booth Unilever Research, Port Sunlight, Wirral L63 3JW, and Department of Psychology, University of Birmingham, Birmingham B15 2TT, England Human responses to odour are acquired in complex settings, by association with emotionally significant effects. Odours thus help to carry the meaning of their sources, evoking recognition and effect. Olfactory discrimination between people involves elicitation of attitude, whether babies' maternal identification or adults' reactions to body odour and perfume. These effects are determined by the odorant's place in a whole stimulus array which may include other odorants, a specific type of social situation and the perceiver's state of mind. The results of an experiment on the social effect of an odour will therefore depend critically on the test situation, procedure and instructions. We use these principles here to resolve apparent conflicts in research findings and to point implications for the formulation and advertising of personal fragrances.

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Social olfaction in human infants: developmental and functional aspects


Benoist Schaal University de Franche-Comti, Laboratoire de Psychophysiologie, 25030 Besanfon, France, and Universiti de Montreal, E.P.E., 750 Bid. Gouin-est, Montreal H2C 1A6, Canada The ethological significance of olfaction has not yet been included in the overall understanding of the infant's psychobiological development. (i) There is little knowledge on the ontogenetic interrelations between structural and functional aspects of human nasal chemosensitivity. Psychophysical experiments using artificial odours have shown that, from the neonatal period, infant's olfactory sensory parameters are nearly similar to that of mature function. However some investigations suggest that the detection thresholds for given odorants appear to be lower and less variable in children than in adults. (ii) During the 1st week after birth, infants are able to rely on this well developed olfactory competence in social contexts, e.g. for individual and kin discrimination. The odour blend collected on the breast-areolar region appears to be an effective cue for reliable discrimination of the mother by the neonate, a prerequisite for the establishment of privileged bonding processes. This presumed olfactory contribution to early infant-mother interaction is not exclusively linked to suckling-related odours since more permanent social odours (e.g. odour mixtures collected on the neck or axillary regions) can be as ethologically salient for the infant. The regulatory effects of maternal odours on infant's behaviour and physiology might thus be perpetuated after the neonatal period. Indeed 3- to 5-year old children are still able to discriminate their mother's or siblings' body odour by smelling a T-short worn by them. (iii) Olfactory based social orientations may be the result of two categories of developmental mechanisms. (a) A pre-natal determination of post-natal olfactory preferences cannot be ruled out a priori: in the fetus, at least three olfactoreceptive structures are present which could extract some information on maternal chemistry from amniotic chemical ecology, and evidence exists of transnatal retention of pre-natal sensory experience. (b) However, present evidence suggests that even if some form of pre-natal influence exists, it is rapidly superseded by post-natal associative mechanisms which work when the infant is exposed to the skin semiochemistry of conspecifics during proximal interactions; for example, breast-fed neonates who have recurrent direct skin-to-skin contact with their mothers are able to discriminate the maternal olfactory characteristics while bottle-fed neonates do not show this ability.

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ECRO VD: Abstracts (iv) To date direct and reproducible studies of the regulatory effects of social odours in early interactions and in the infants' psychophysiological balance are still rare. However clinical evidences underline: (a) their potential implication in the regulation of the infant's behaviour during nursing (odours can provide qualitative information about the nature of ingested substances and can act as incentives by their regular pairing with suckling episodes); (b) that odours of familiar individuals or olfactory self-perception appear to be significant dimensions of the infant's emotional homeostasis, especially during separation episodes from the attachment figures; (c) that body odours are involved in the child's attraction or aversion behaviours to other individuals.

V. Oral Communications
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1. Olfaction and Trigeminal Chemoreception Axonal contribution to olfactory receptor cell adaptation
G.D.Adamek and R.C.Gcsteland Department ofAnatomy and Cell Biology, University of Cincinnati Medical Center, Cincinnati, OH 45267, USA The olfactory nerve in frogs is comprised of unmyelinated axons of uniform diameter of 0.2 /im. They lie in groups of hundreds or thousands within invaginations of Schwann cells. This arrangement is unique among sensory neurons. At appropriate concentrations, odors commonly evoke action potential trains from individual cells with rates of up to 30 Hz. Electrical stimulation of the olfactory nerve with bipolar hook electrodes at 1 Hz shows a very different activity pattern. A cell which displays a time-locked action potential in response to anti-dromic stimulation responds to the first 56 shocks, then fails a few times and is therefore mostly unresponsive. It is conceivable that anti-dromic and orthodromic conduction processes are different. However, an electrical stimulus delivered to the synaptic terminals of the axons by placing a bipolar electrode in the olfactory bulb, shows this to be untrue. Cells can respond faithfully to each shock at frequencies > 10 Hz for many seconds. Following repetitive stimulation at 10 Hz, the theshold value for anti-dromic stimulation increases only slightly. The latency varies between 70 and 90 ms, depending on the placement of the electrode in the epithelium. As expected, the latency of spikes recorded from areas more caudal in the epithelium was less than from spikes recorded more rostrally. The crucial difference between these experiments is that bulbar electrical stimulation and odor stimulation activate only a few neurons, while nerve stimulation excites most of the 50 million or so axons synchronously. In the latter case potassium concentration in the extra-axonal space must increase and block conduction via conductance inactivation. Thus, signalling properties of the olfactory nerve are not well revealed by studies of the compound action potential. Responses to natural stimuli which activate a large fraction of the cell population must adapt quickly. The anatomical arrangements of the olfactory nerve acts as filter to accentuate the effectiveness of natural stimuli which activate cells whose axons are not in close proximity. Supported by Grants NIH PO1 NS18490-04-08, NIH RO1 NS23523-O1-O7 and NSF BNS-85^14025.

Neurogenesis in the olfactory epithelium


W.Breipohl, A.Mackay-Sim1, B.Rehn1 and D.Walker University of Queensland, ^University of Adelaide, Australia, and 'University of Essen, FRG Evidence is given for age-related changes in olfactory receptor cell maturation in post-natal and adult mice which extend previous investigations on the neuronal plasticity in the olfactory system (Breipohl and Ohyama, 1981; Breipohl el al., 1985, 1986; Mackay-Sim and Patel, 1984; Rehn and Breipohl, 1986; Rehn et al., 1986). The data challenge the concept of a regular neuronal cell turn-over in the olfactory epithelium due to an inherent clock of the sensory cells. Parameters investigated include age-related histological changes in the epithelial compartments, mitotic activity, regenerative capacity, ratio of progenitor cells versus receptor cells, cell death phenomena in the receptor cell compartment as well as in the progenitor cell compartment, regional shifts of these parameters within the same animal. First estimations on the relative percentage of mature and almost mature olfactory receptor cells will be presented as further evidence against the general idea of a regular cell turnover. On the basis of these data a new hypothesis on neuronal cell plasticity is proposed, which assumes that

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ECRO VII: Abstracts at least some of the olfactory receptor cells tend to live for as long as possible provided they are not killed by environmental insults or internal factors. BreipohJ.W. and Ohyama.M. (1981) Biomed. Res., 2 (Suppl.), 437-448. BreipohJ.W., Rehn.B., Molyneux.G.S. and Grandt.D. (1985) XII International Anat. Congress, London (Abstract). Breipohl.W., Mackay-Sim.A., Grandt.D., Rehn,B. and Darrlemann.C. (1986) In BreipohJ.W. and Apfelbach.R. (eds), Principles of Maturation of the Olfactory System of Vertebrates. Springer, Berlin, in press. Mackay-Sim.A. and Patel.U. (1984) Exp. Brain Res., 57, 99-106. Rehn.B., Breipohl.W., Memdoza.A. and Apfelbach.R. (1986) Brain Res., in press. Rehn.B. and Breipohl.W. (1986) In Breipohl.W. and Apfelbach.R. (eds), Principles of Maturation of the Olfactory System of Vertebrates. Springer, Berlin, in press. Supported by grants: ABF 9295, BRF 52, DFG-Br. 358/5-2, NH&MRC 860587:85/3452 and 850400:84/4993. Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on February 16, 2014

The effects of tongue movements and swallowing on retronasal aroma perception


Konrad J.Burdach and Richard L.Doty University MOnchen, D-8O00M0nchen, FRG and University of Pennsylvania, Philadelphia, PA 19104, USA Everyday sensory experience suggests that the aroma intensity of food substances may increase considerably during mastication and, especially, during swallowing. The present study was designed to analyse this sensory phenomenon empirically. In a repeated measurement setting eight subjects assessed the aroma intensity of two stimulus substances (rum and orange aroma diluted in deionized water) under various experimental conditions, particularly after (standardized) tongue movements, after expectorating the stimulus substance and after swallowing it. The subjects rated the aroma impression by pulling out a tape-measure according to the perceived stimulus intensity. The measurement procedure was repeated four times. The results are based on a comparison of the medians of the (Jog-transformed) magnitude estimations in the different experimental conditions. They show that, compared to a control condition (no movements), tongue movements and swallowing increased the perceived aroma intensity dramatically. An unexpected finding was that the expectoration of the stimulus substances produced nearly the same (aroma enhancing) effect as swallowing. These results emphasize the dynamic nature of retronasal aroma perception.

Quantitative determination of the volatiles produced by living organisms


B.V.Burger and Z.M.Munro Laboratory for Ecological Chemistry, University of Stcllenbosch, Stellenbosch 7600, South Africa A system has been devised with which volatiles can be quantitatively trapped from gas samples at relatively high flow-rates. Material is trapped in fused silica traps, 1 m in length and coated with either an immobilized stationary phase layer, or a suitable adsorbent such as activated carbon or a powdered porous organic polymer such as PorapaJc Q supported on immobilized SE-30. Thercmal conversion of labile compounds is avoided by temperature-programmed desorption in conjugation with cold trapping of the desorbed volatiles on the capillary column. The versatility of this headspace-analytical technique for the determination of air-borne volatiles produced by living organisms was demonstrated by the determination of the quantitative composition of the multicomponent sex-attracting secretions of individual males from two insect species. The difference in the composition of the material released into the air and that of material extracted from the pheromone-producing glands of these insects is discussed. This technique can also be effectively used for the determination of the headspace volatiles of, for example, wine and other beverages.

The modified glomerular region and pattern of 2-DG uptake in the olfactory bulb of rabbit pups exposed to nipple-search pheromone
H.Distel, H.Martin and R.Hudson Institute of Medical Psychology, University of Munchen, D-S000 MOnchen, FRG Rabbit pups are only nursed for - 3 min once a day. To locate nipples in this short time they are totally dependent on a short ranging pheromone on the mother's belly which releases and guides a highJy stereotyped

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ECRO VII: Abstracts nipple-search behaviour. This is the case even for pups delivered by caesarian section, and thus without post-natal experience of the pheromone. Newborn rats also depend on odour cues to locate nipples, and 2-deoxyglucose (2-DG) studies of activity in the olfactory bulb have indicated a modified glomerular region adjacent to the accessory bulb to mediate these cues [Teicher et al. (1980) Brain Res., 194, 530]. It was thus the purpose of this study to examine whether a similar region is present in the newborn rabbit and, using the 2-DG method, whether it is specifically activated during exposure to the pheromone. Five-day-old rabbit pups were injected intracardially with 300 /xCi of 14C-labelled 2-DG and then placed for 45 min on either the belly of an anesthetized, pheromone-producing female rabbit, on a non-pheromone producing female or on a non-pheromone producing female scented with the odour of citral, to which pups had been conditioned to respond with nipple-search behaviour [Hudson (1985) Dev. Psychobiol., 18, 575]. The autoradiograms revealed loci of high 2-DG uptake adjacent to the unlabeled accessory olfactory bulb, which did indeed correspond, when compared to the Nissl sections, to a morphologically distinct region. In the rabbit, this region appears to form a crescent around the anterior nm of the accessory olfactory bulb and is characterized by a fusing of glomeruli, a much reduced external plexiform layer and the loose arrangement of the numerous mitral cells below. However, 2-DG labeling of this 'modified glomerular region' was not confined to animals exposed to nipple-search pheromone but was also clearly seen in both control conditions. Further, exposure to the pheromone resulted in patches of strong labeling distributed throughout the glomerular layer of the main bulb. A similar but much weaker pattern of activity was obtained for the non-pheromone controls, whereas a strong but not so widespread labeling of glomeruli was observed in the citral condition. Thus, although pheromone exposure resulted in the strongest labeling, the actual pattern of activity could not be regarded as distinctive, particularly with respect to activation of the modified glomerular region. Supported by the Deutsche Forschungsgemeinschaft, Di 212/3-1.

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X-Irradiation of embryonic and newborn rats: effects on olfactory bulb structure


N.Halasz Institute of Biophysics, Biological Research Centre, Hungarian Academy of Science, Szeged, Hungary Neurons are most susceptible to X-rays during migration and proliferation, while mature neurons are relatively resistant to low-dose irradiation. We have tested various doses of X-rays in order to find the appropriate magnitude for damaging specific classes of neurons in the rat olfactory bulb (OB). Total doses between 210 and 1070 rads were fragmented into 50-200 rad single doses and delivered according to different schedules between day 8 of embryonic life (in utero irradiation) and post-natal day 12. Dams and young pups were characterized for weight gain and mortality and young rats were studied to reveal morphological changes in the OB, by light and electron microscopy. In general, pregnant mothers as well as pups were retarded in their development by the irradiation. The OBs were considerably smaller than those of the controls and in several experiments, development of the granule cells, granule cell layer, external plexiform and glomerular layers were also far behind that of normals. The observed changes were proportional to the magnitude of irradiation in most cases. As a summary, the perinatal irradiation technique seems to be useful in damaging certain, even pre-natally developing, classes of the OB neurons.

Regional autonomy in the peripheral processes of nipple-search odors


R.Hudson and H.Distel Institute of Medical Psychology, University of MUnchen, D-8000 Munchen, FRC Newborn rabbits are completely dependent on pheromonal cues on the mother's belly for the release of nipplesearch behavior, but can be rapidly conditioned to respond with the same search behavior to other odors when these are paired with suckling [Hudson (1985) Dev. Psychobiol., 18, 575]. As reported in the accompanying abstract (No. 5), 2-deoxyglucose (2-DG) labeling is found throughout the olfactory bulb following exposure to such nipple-search odors. This raises the question whether the entire bulb is necessary for mediating these and, if not, whether specific areas, such as the modified glomerular region, are essential. To test this question, extensive bilateral lesions of the medial, lateral or dorsal bulb were performed on day 4 after pups had been conditioned to the odor of citral or isopropyl alcohol. The next day pups' responsiveness to both the pheromone and the conditioned odor was tested, as well as their ability to obtain milk

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ECRO VII: Abstracts in the normal nursing situation in competition with unoperated pups. Lcsioned pups were able to suckle in the nest, and searched vigorously on a test female scented with the conditioned odor, even after one nare had been temporarily closed. Only in a few severely lesioned cases was a decrement in nipple-search performance observed. Similar lesions were also performed on naive pups 2 h after birth. They, too, could respond to the pheromone and suclde when tested the following day, and furthermore, could be conditioned to the novel odors. Reconstruction of the lesion sites showed them to overlap fully so that no part of the bulb had been completely spared, and that even pups with <30% of the bulb remaining intact could still respond with search behavior to the various odors. These results not only confirm the earlier finding that the veromonasal organ is unimportant for the mediation of such nipple-search odors [Hudson and Distel (1986) Physiol. Behav., 37, in press], but also show the modified glomerular region not to be essential. In fact it would seem that any part of the main olfactory bulb of newborn rabbits is capable of autonomously processing the pheromone or a post-natally acquired artificial odor. By implication, this also seems to be true for the olfactory epithelium as this is known to project in a broadly topological manner to the bulb. However, whether these findings hold for all odors, and for the adult animal remains to be investigated. Supported by the Deutsche Forschungsgemeinschaft, Di 212/3-1.

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Olfactory and chemo-somatosensory evoked potentials from stimuli presented to the left and right nostrils
G.Kobal, Th.Hummel and C.Van Toller1 Department of Pharmacology and Toxicology, University of Erlangen-NUrnberg, Universitatsstr. 22, D-8520 Erlangen, FRG, and ^Department of Psychology, University of Warwick, Coventry CV47AL, UK The aim of the present study was to determine whether topographical differences of olfactory and chemo-somatosensory evoked potentials can be observed in relation to the stimulated nostrils. Fourteen subjects participated in the experiments. The EEG was recorded from Fz, Cz, Pz, C 3 , C 4) F 3 , F 4 and Fp, referenced to A, according to the international 10/20 system. The stimulation device produced stimuli of 200 ms duration (CO2, menthol, HjS, vanillin) without tactile somatosensory contamination. The stimulus interval was 4050 s. Four separate classes (n = 10... 16) were used to average the evoked potentials: right stimulus (A), left stimulus (B); correct subjective identification of stimulated side (C), incorrect subjective identification of stimulated side (D). Maximal amplitudes to CO2 and menthol were in Cz and decreased conspicuously more to the frontal than to the parietal part of the skull. In general they were larger in the right hemisphere than in the left, independent of the side actually stimulated. Earlier potential components appeared first centrally and parietally, and then frontally, whereas later components had the opposite order of appearance. Stimulation of the left side occasioned shorter latencies. Responses to HjS and vanillin presented a different topographical aspect. Their maximal amplitudes were located parietally, but depended on the stimulated side. When HjS was used as stimulant, the earlier potential components were largest after stimulation of the right side; with vanillin the largest responses, and a simultaneous elongation of the latencies, appeared after stimulation of the left side. In cases where the subjective identification of side was correct, stimulation with HjS and vanillin yielded shorter latencies, in cases of incorrect identification evoked potentials appeared later and were larger, although subjects made correct and incorrect decisions of the same frequency. This experiment established the utility of evoked potentials to distinguish between the quality of odours and between cognitive situations.

A comparison of cytochrome oxidase activity and 2DG labelling in the rat olfactory bulb
H.Panhuber, G.A.Bell and D.G.Laing CSIRO Division of Food Research, PO Box 52, North Ryde, NSW 2113, Australia Both cytochrome oxidase activity and glucose uptake (as measured by [14C]2-deoxyglucose-labelling) can indicate differences of metabolic activity levels within neurological tissue. Cytochromes are energy-deriving enzymes responsible for electron transport and oxidative phosphorylation yielding ATP (adenosine triphosphate), they are most active on the inner membranes of mitochondria. 14C-Labelled 2-deoxyglucose (2DG) is taken up in competition with glucose in energy-consuming regions of the brain but is not completely metabolised and remains in the tissue.

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ECRO VD: Abstracts The two techniques however, mark different parts of the olfactory bulb. The intensity of the marking varies in different ways with the two techniques according to the stimulus conditions to which rats have been exposed. Cytochrome oxidase activity appears to be greater in the external plexiform layer than in other parts of the bulb, while levels of 2DG label are generally low in this layer and are higher in the glomerular and granule cell layers. The issue is to show how metabolic activity in the external plexiform and glomerular layer, as shown by cytochrome oxidase, relates to the activity shown in the glomerular layer by 2DG.

Morphometric study of the glomerular population in the mouse olfactory bulb: numerical density and distribution of the size along the rostrocaudal axis
J.P.Royet, F.Jourdan, C.Souchier1 and H.Ploye2 Laboratoire de Physiologic Neurosensorielle, ICBMC, F-69622 Villeurbanne Cedex, ''Centre Commun de Quantimttrie, 8 avenue Rockefeller, 69373 Lyon, and 'Laboratoire de Biomitrie, IASBSE, F-69622 Villeurbanne, France It has been shown that the size of the mitral cells varies around the coronal but not along the rostrocaudal extent of the olfactory bulb (Panhuber et al., 1985). These patterns of size variation may be due to a functional differentiation possibly associated with the amount of information received by each mitral cell and/or with differences in their glomerular arborisations. The aim of this study was to determine whether the size and distribution of glomeruli in the olfactory bulb of the mouse are uniform or not. Serial histological sections, 20 /im thick, were studied with an image analyser (Quantimet 900, Cambridge Instruments). In order to avoid several measurements of the same glomerulus, one section in 10 was analysed. Thus, 13 sections were studied in each olfactory bulb. All the glomeruli were accurately delineated with the light pen and their individual area measured. Afterwards, the whole glomerular layer was also delineated and its area computerized. Glomeruli were considered as spheroids (convex objects). Therefore, we applied the stereological melhod recently developed by Cruz-Orive (1983). This method takes into account the problems of the estimation of the small size profiles and of the section thickness. The results show that the diameters of 91 % of the glomerular range from 60 to 130 /un. Moreover, the estimation of the mean caliper diameter (averaged for orientation) computerized for eight mice shows that the largest glomeruli are located at the middle of the rostrocaudal axis. Since the surface density of the glomeruli remains constant along the rostrocaudal extent of the olfactory bulb (average: 0.56), it is not surprising to observe that the smallest numerical density is also found at the middle of the olfactory bulb. Finally, the computerization of all these data led us to estimate that the number of glomeruli is ~ 1400 within the olfactory bulb of the mouse. Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on February 16, 2014

Quantitative study of the glomerular activation patterns in the mouse olfactory bulb: densitometrk analysis of autoradiograms labelled with the P^CJI-deoxyglucose
J.P.Royet, G.Sicard, C.Souchier1 and F.Jourdan Laboratoire de Physiologic Neurosensorielle, ICBMC, Universitt Claude-Bernard, F-69622 Villeurbanne, and ^Centre Commun de Quanlimitrie, 8 avenue Rockefeller, F-69373 Lyon, France The purpose of this work was to examine the spatial distribution of the activity evoked by stimulations in the glomerular layer of the mouse olfactory bulb. The mice were stimulated with either an odorous stimulus (amyl acetate or isovaleric acid) or pure air. The autoradiographic method based on the use of the [14C]2-deoxyglucose was applied to measure the functional activity of the olfactory glomeruli. The serial histological sections of the olfactory bulb and the corresponding autoradiograms were analysed with an image analyser, the Quantimet 900 of Cambridge Instruments. In order to avoid measuring die functional activity from the same glomerulus several times, one section in 10 was analysed. Therefore, 13 sections were studied in an olfactory bulb. The outlines of the histological sections and of the corresponding glomerular layer were accurately delineated. They were superimposed with the autoradiographic films and the optical density along the glomerular layer was automatically measured with 13 sections being studied in one olfactory bulb, we obtained 13 OD profiles allowing us to establish a glomerular activation map. To test the reliability of the measurements, one subject was chosen at random in each mouse group and analysed twice. To control the symmetry of the glomerular labeling in the two bilateral olfactory bulbs, analysis of the contralateral olfactory bulb was performed in three subjects. A principal component analysis of the olfactory maps obtained from the mice stimulated with different odours shows that the glomerular activation patterns are characteristic of the odorant stimulus.

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Mitral cell response patterns revisited


D.Schild Physiologisches Institut der Universitat Humboldxallee 23, D-3400 Gtittingcn, FRG Mitral cells of the olfactory bulb respond with patterned discharges to natural stimuli. In quantitative analysis of experimental data, the temporal structure of responses has rarely been taken into account: there are only two attempts to classify mitral cell responses (Kauer, 1974; Meredith and Moulton, 1978). In this study wereinvestigatethe response pattern problem taking into consideration two facts: (i) the stochastic fluctuations of mitral cell activity are large, i.e. many stimulus responses have to be superimposed in order to eliminate noisy signal components to an acceptable degree; (ii) responses that are not always reproducible (Schild and Zippel, 1986), should not be considered when pattern features are defined because unreproducible responses should not be regarded as drawn from the same (multidimensional) distribution. For every stimulus we recorded 40 responses (with at least 30 min interstimulus interval) and discarded experiments exhibiting unreproducible responses. Having 40 activity values in each bin allows the statistical comparison (U-test) of the activity in a particular bin A to the activity of another bin B. When this kind of activity comparision is done for all combinations of bins a very simple result emerges: at the beginning of a response there is a dynamic response component during which the activities per bin do not differ from each other. This component is followed by a static component; the activities per bin during this second component also do not differ from each other. In conclusion, there are only two pattern features of mitral cell responses: the mean activities of the dynamic and the static response components. Responses thus have one of the following nine types: dynamic response inhibitory ^^ZZZZ^Z_ static response inhibitory indifferent excitatory inhibitory indifferent excitatory inhibitory indifferent excitatory

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indifferent

srZZZZ

excitatory

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Nasal trigeminal chemoreception: responses to aliphatic acids


Wayne L.Silver and Dianne Walker Department of Biology, Wake Forest University, Winston-Salem, NC 27109, USA Trigeminal receptors in the nasal cavity respond to many volatile odorants. However, the properties of odorant molecules which make them effective trigeminal stimuli are largely unknown. Previously [Silver el al. (1986) Brain Res.], we demonstrated that sirmulus effectiveness increased with increasing carbon chain length for all homologous series of aliphatic alcohols. We speculated that lipid solubility might be an important factor in determining the effectiveness of trigeminal stimuli. In the present experiment, integrated multiunit recordings were obtained from the ethmoid branch of the rat trigeminal nerve in response to eight n-aliphatic acids (formic, acetic, propionic, butyric, valeric, hexanoic, heptanoic and octanoic) to test further this hypothesis. In addition, respiratory rate was monitored by means of a thermocouple wire placed in a trachea! cannula. Stimuli were presented via an air dilution olfactometer, at a flow-rate of 1 1/min. Air was drawn through the nasal cavity via a nasopharyngeal cannula at a rate of 0.2 l/min. Multiunit responses were obtained to all eight acids, although only high concentrations of the lower mol. wt compounds (formic acid, butyric acid) reduced the rat's respiratory rate. All rats did not respond to C,C1. Thresholds for the eight acids, defined as the concentration which first elicited a multiunit response greater than the baseline activity, decreased with increasing carbon chain length, with the exception of valeric acid. Since lipid solubility in these acids increases with increasing carbon chain length, an inverse relationship between lipophilicity and stimulus effectiveness is again seen. An explanation for these findings is that the more lipophillic a substance, the more easily it can penetrate mucous and epithelial layers to reach chemosensitive trigeminal nerve endings. Supported by NIH grant no. NS-23326.

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Odour localization and psychometric results from olfactory and trigeminal stimuli presented to the left and right nostrils
C.Van Toller, Th.Hummel1 and G.Kobal1 Department of Psychology, University of Warwick, Coventry CV4 7AL, UK, and ^Department of Pharmacology and Toxicology, University of Erlangen-NUmberg, UniversitOtsstr. 22, D-8520 Erlangen, FRG In 1964 von Bekesy stated that the human sense of smell possesses an equivalent to the phenomenon of directional hearing in the auditory system. Contradictory to this assumption, older and more recent publications denied odour localization mediated by the olfactory nerves. The aim of the present study was to establish the crucial pre-condition for a neuronal processing of local olfactory information, that is the ability of humans to discriminate between odorous stimuli perceived either from the right or from the left side. In our study an olfactometer that allowed stimulation of each nostril separately, randomly applied stimuli to both sides, using a computer monitor the subjects were asked to indicate the side on which they had perceived the stimulus, and to state how sure they were of this decision. In separate experimental sessions the substances HiS, vanillin, menthol and CO2 were presented 30-40 times each. Fourteen subjects participated in the experiments. The correct identificaton rates were 47.0 and 44.5% when vanillin or HjS were used as stimulants, which indicates that the localization of both substances was random. On the other hand stimulation with menthol or CO2 yielded identification rates of 96.6 and 97.4%. These results established the fact that directional orientation, considering single momentary odorous sensations, can only be assumed, when the olfactory stimulants simultaneously excite the trigeminal somatosensory system. This research was supported by the Sander-Stiftung and by a twinning grant of the European Science Foundation.

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Ontogenetic analysis of the rat pup pheromone implicated in perigenital licking


Evelyne Vernet-Maury, Isabelle Brouette-Lahlou and Jacques Chanel Laboratoire de Physiologie neurosensorielle, CNRS, University Claude Bernard/Lyon, F-69622 Vtlleurbanne Cedex, France In the rat, perigenital licking of the pups by the mother is vital during the first 3 weeks of their life. Young male rats are licked longer than females (28 and 22 days, respectively). We pointed out that secretions of the pups preputial glands allow the mother to identify licked pups from non-licked ones and regulate licking time. Two principal compounds were isolated from these secretions by gas chromatography and mass spectrum anlaysis: diethylhexyl adipate and dodecyl propionate. They were synthesized. The latter only induced licking behavior. We considered it as a pheromone. From this point of view it as important to determine the onset and evolution of these two compounds during the pups life and according to their sex. In the present experiments, we point out that dodecyl propionate (chromatographic peak) was present as early as birth, and became more and more important during the first 9 days. It remained constant in the males until adulthood but diminished regularly in the females and was no longer present after the 25th post-natal day. Diethylhexyl adipate can be detected only the 17th day and it becomes more and more important afterwards, especially in female pups. Different new peaks appear on the 35th post-natal day: they characterize adulthood. This ontogenic analysis confirms dodecyl propionate as the pheromone of perigenital licking; it may also explain why male pups are licked longer than females.

Functional recovery and morphological reconstruction in the goldfish olfactory system after lesions in peripheral and central structures
H.P.Zippel and D.L.Meyer Physiologisches Institut, Humboldtallee 23 and institutfUr Anatomic, Kreuzbergring 36, Universitat, D-3400 GCmngen, FRG A special aspect of neuronal plasticity, the regeneration of CNS pathways and nuclei, is little understood in its basic aspects. The olfactory system became due to its enormous peripheral regenerative capacity of increasing interest for the study of regeneration and the underlying principles. The research in this field was mainly focused on the olfactory mucosa (receptor de- and regeneration, and EOG changes following receptor axotomy). The present investigations were made to demonstrate the correlation between morphological restitution in the olfactory mucosa and in the olfactory bulb, with behavioural observations after

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ECRO VH: Abstracts dissection of different parts in the peripheral and central olfactory system. For this purpose the pre-operative spontaneous and training behaviour was compared with the post-operative reactions of the same animals. Immediately following surgery the animals fail to respond to low concentration odour stimuli but readily react to stimuli above the taste threshold. A return to the pre-operative behaviour could be recorded at different time intervals post-operatively, and is in accordance with the neuro-anatomical findings [HRP studies for demonstration of the connections between the olfactory mucosa (MO) and the olfactory bulb (BO), and light microscopy for investigation of the olfactory receptors in the MO]. After bilateral receptor axotomy [see inserted sketch (1)] or following bilateral extirpation of the rostral halfs of the BO (4) the fish respond positively after an extremely short regeneration period of - 7 - 1 0 days, whereas after bilateral dissection of the olfactory tracts (2) or extirpation of the caudal halfs of the bulbs (5) the animals return to the preoperative behaviour after ~ 6 weeks. Following bilateral total bulbectomy (3) the fish do not respond to low threshold stimuli even after a 12 month regeneration period (Zippel et al., 1981); in this case the receptors in the MO regenerate, their axons form a neuroma at the place of the former BO, but no connections could be found between the olfactory mucosa and the telencephalon.

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Mucosa olfactoria Fila olfactoria


Bulbus olfactorius Tractus olfactorius medialis et lateralis

2. Taste

Multisapophoric molecules
G.G.Birch University of Reading, Reading RG6 2AP, UK Molecules possessing two or more sapophores may exhibit one or more basic tastes. The reason for this phenomenon is not clear from simple structureactivity relationships and may be due to solutewater interactions. This communication examines some typical multisapophoric molecules and their solution properties in an attempt to explain how the solute may be disturbing water structure. Apparent molar volume and intrinsic viscosity are two properties which allow the interaction of solute molecule with water structure to be investigated. The different regions of each solute molecule interact differently with water molecules and this gives rise to clustering of water molecules around solute molecules. Some of these effects could account for the observed tastes of multisapophoric molecules and could lead to an explanation of the mechanism of taste chemoreception.

Relationship between the suppressive effect of different sweet substances


J.H.A.Kroeze
Laboratory of Psychology, University of Utrecht, Sorbonnelaan 16, NL-3584 CA Utrecht, The Netherlands Taste mixtures of different quality show mutual suppression. This can be assessed by applying magnitude estimation of the quality of each mixture component. However, when the components have identical quality, it is impossible to estimate them separately. With sucrose and saccharine, there is no direct way of separating both sweetnesses. Therefore, we tried a different way and applied it to the problem of common versus different receptor sites for sweet substances. Saccharine and sucrose, were used. Each of the substances was mixed with a third substance of different taste quality. The subjective intensity of this third substance was then assessed. Sucrose was mixed with quinine in one group of trials and saccharine was mixed with quinine

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ECRO VII: Abstracts in another group of trials. The subjects estimated bitterness. In this way we got two separate types of bitterness suppression, one type caused by saccharine, the other caused by sucrose. The crucial question is: are these two types of suppression related? If they appear to be unrelated, we must reject the idea of a common receptor site for the two sweeteners under investigation. The experiment was repeated with citric acid as a test substance. Moreover, the conditions were divided into 'mixtures' of which the components were separated by the medial plane of the tongue versus mixtures where the components were mixed on the tongue. In both experiments saccharine and sucrose showed essentially different results: there appeared to be no relationship between the way in which they suppressed the test substance. Suppression of the test substance by saccharine cannot be predicted from suppression by sucrose. Results do not support the hypothesis of a common receptor site for both sweeteners.

Linking the biophysics and psychophysics of taste


Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on February 16, 2014 Robert L.McBride CS1RO Division of Food Research, PO Box 52, North Ryde 2113, Australia When the taste response of animals is measured neurophysiologically, the relationship between response and (log) stimulus concentration conforms to a predictable, sigmoidal curve. This relationship is commonly described by the Beidler equation, RJRmlx = CK/(l+CK), where R is the neurophysiological response to stimulus concentration, C; ^ ^ is the maximum (saturated) response to the stimulus; and K the association constant of the stimulus. Recent work shows that the stimulus/response relationship for human taste, as determined by two different psychophysical methods (JND cumulation, category rating), also conforms to the sigmoidal curve, consistent with the Beidler equation. Thus, the human taste sensation may simply and directly reflect the underlying neurophysiological response. Application of the Beidler equation to psychophysical data has implications for both basic and applied research. On the basic side, determination of K and R^^ provides some insight into stimulus biophysics, hitherto not available from human psychophysical data. On the applied side, the Beidler equation opens the way to express taste intensities in terms of a 'Taste Coefficient' (RIRTrm). The expression of taste intensities in terms of a universal Taste Coefficient would render the measurement of taste less idiosyncratic.

Lectin histochemical characteristics of taste bud mucous substances in different vertebrates


K.Reutter and M.Witt Anatomical Institute, University of Tubingen, FRG The taste buds (TB) of vertebrates even belonging to different systematic groups and to different living species, are always covered by mucous substances. In different species the thickness of this mucous layer, its viscosity and also its chemical structure vary. By using conventional carbohydrate histochemical methods the mucous is to be characterized as containing variable amounts of sialomucins and sulfomucins (1). As it is suggested that these mucous substances have not only protective functions at the TBs receptor field, but also functions regarding chemoreception (2,3), we continued to characterize and to compare TB mucous subtances by using the lectin-binding methods. Lectins possess a great affinity for terminally positioned monosaccharides and oligosaccharides. The determination of such sugars seems to be essential in view to the question whether the TB mucus is able to fulfil receptor-like functions or not. The investigation was done on the TBs of a fish (Silurus glanis), a frog (Rana temporaria), a bird (Commix japonica) and the rabbit. We applied the avidin-biotin/peroxidase complex (ABC) method (4) and visualized the reaction product with diaminobenzidine (5). As a result it is to be stated that the mucous substances covering the fish's TB possess all the terminal positioned sugars for which we searched. In contrast, the mucous layer of the frog's, bird's and the rabbit's TBs show a more differentiated mode of terminal sugar representation, e.g. a-L-fucosc and the disaccharide /3-D-Gal-(l-3)-D-GalNAc are lacking. These differences possibly indicate that in animals of different systematic position the mucous substances of the TBs are closely related to special and species-specific functions of these organs. (1) Reutter,K. and Klessen.Ch. (1979) Verh. Anal. Ges., 73, 10189-1021. (2) Bannister,L.H. (1974) In Poynder,T.M. (ed.), Transduction Mechanisms in Chemoreception. Information Retrieval Limited, London, pp. 3946.

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ECRO VII: Abstracts (3) Reuter.K. (1980) In van der Starre.H. (ed.), Olfaction and Taste VII. 1RL press, London, and Washington, p. 107. (4) Hsu.S.M. and Raine.L. (1982) /. Histochem. Cytochem., 30, 157-161. (5) Graham.R.C. and Karnovsky.M.J. (1966) J. Histochem. Cytochem., 14, 291-302.

Recent findings on transduction and modulation of taste


S.S.Schiffman, T.G.Beeker and Z.S.Warwick Department of Psychiatry, Duke Medical Center, Durham, NC 27710, USA In the past several years, new insights into the mechanisms of taste transduction and modulation have been achieved by observing the psychophysical and neurophysiological changes that occur after application of pharmacological agents to the surface of the tongue. The effects of five compounds on the taste system will be discussed; they are amilonde, caffeine (and other methyl xanthines), inosine (and inosine monophosphate), kainic acid and BaCl2. Amilonde, a potassium-sparing diuretic that has been used extensively as a specific inhibitor of sodium transport, has been found to inhibit the taste of sodium salts and sweeteners. Caffeine and other methyl xanthines enhanced the taste of artificial sweeteners (acesulfam-K, neohesperidin dihydrochalcone, d-tryptophan, thaumatin, stevioside and sodium saccharin), NaCl and quinine HC1. Methyl xanthines at the concentrations used inhibit Aj adenosine receptors. Inosine (and inosine monophosphate) potentiated the taste of sucrose and aspartame. Kainic acid, a glutamate agonist, blocked the taste of glutamic acid in rats. Barium chloride blocked the taste of KC1. These data suggest that sodium and potassium channels, as well as receptors for adenosine and inosine, play an important role in taste perception.

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Sweet taste intensity and taste similarity of mixtures containing neohesperidin dihydrochalcone (NHD) with either sucrose or saccharin
Lyat Yaron, Michael Nairn, Uri Zehavi and Martha Levinson Department of Biochemistry and Human Nutrition, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76-100, Israel The slope values of sweet taste intensity curves of mixtures containing NHD with either sucrose or sodium saccharin and taste similarity experiments were used to evaluate the sweet quality of these mixtures. Slope values of sweet intensity curves of mixtures containing NHD and saccharin were not different from those of NHD alone. Mixtures containing NHD and sucrose, however, in which NHD contributed 40% and 60% of total sweet intensity produced slope values significantly higher than that of NHD. Both, the three-dimensional scaling (MDS) and the hierarchical tree structure (HTS) expression of taste similarity suggested that the taste quality of a mixture of NHD and sucrose in which NHD contributed 40% of total sweet intensity was very similar to that of sucrose. No such improvement, however, was found for the respective mixture of NHD and saccharin. All mixtures containing higher proportions of NHD (e.g. 60% and 80%) were located further than that containing 40% NHD in relation to the sugar area. The present results support the suggestion that an increased slope value of the sweet taste intensity curve of a mixture is one indication for improved sweet quality.

3. Chemoreception in Invertebrates

Chemoreceptor adaptation: a patch in space is a pulse in time


Jelle Atema Boston University Marine Program, Marine Biological Laboratory, Woods Hole, MA 02543, USA Recent results obtained from studies on the spectral and dynamic filtering properties of receptor cells in the lobster, Homarus americanus, having shown that these cells self-adapt and - despite narrow tuning cross-adapt. Recovery from self-adaptation can take several tens of seconds. Tuning and cross-adaptation studies lead to a receptor cell model in which the cells of a tuned population are characterized by a predominance of one type of molecular receptor for the best compound and an unpredictable variety of different molecular receptors for second, third, etc. best compounds. The results are consistent with the notion that all receptors have independent access to the molecular excitation and adaptation mechanisms of the cell. This model needs to be tested with intracellular methods.

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ECRO VD: Abstracts Data from physical, chemical and biological oceanography combined with the results from self- and crossadaptation studies lead to the hypothesis that receptor cells under natural stimulus conditions must function in some state of adaptation almost continuously. This would be similar to what is known from vision and hearing research. We measured temporal patterns evoked continuously in a stationary detector by the spatial stimulus distributions in a moving aquatic odor plume. The sampling volume of the detector was matched to that of a lobster nose. The pulsed appearance of the resulting signal was very similar to the temporal patterns obtained in aenal odor plumes (Murlis and Jones, 1981). In addition, many animals, including the lobster, employ pulsed sampling behavior, i.e. sniffing or flicking. I propose (i) that one of the functions of receptor adaptation is to create a temporal sampling window for relevant odor pulse frequencies, and (ii) that the animal infers spatial, typically patchy, stimulus distributions from temporal sampling. Different animals, and different chemoreceptor organs in the same animal, should have receptor cells with different adaptation time courses, each one designed to match dominant/relevant stimulus frequencies in its microenvironment (Atema, 1985, 1986). Similar reasoning can be applied to animal behavior designed to interface with spatial odor distributions. Thus, spatial distributions of chemical stimuli are sampled through temporal resolution filters. Supported by the Whitehall Foundation and NSF BNS 85-12585.

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Self-adaptation and cross-adaptation in ammonium receptor cells


Paola F.Borroni and Jelle Atema Boston University Marine Program, Marine Biological Laboratory, Woods Hole, MA 02543, USA In this study we provide the first systematic description of the effects of self- and cross-adaptation in primary chemoreceptor cells. Responses of narrowly tuned NH4-sensitive cells were recorded extracellularly from taste organs of the lobster, Homarus americanus. Single cell stimulus-response functions (0.003-30 mM NHiCl) were recorded in five different self-adapting background concentrations (0.00163 mM NH,C1); the mean stimulus-response functions resulting from these data form a family of five parallel curves, each closely fitted by a logarithmic equation (r = 0.96-0.98). Self-adaptation of the receptors reduced the magnitude of the mean response to all stimulus concentrations by a nearly constant amount (45 spikes/1 log unit of background concentration); similarity in the slope of the mean functions in each background suggests that the kinetics of stimulus-receptor interactions are not changed in the adapted receptor cells. Approximately 40% of all NH4 receptor cells tested do not respond to compounds other than NH4C1; some NH, cells respond weakly to other nitrogenous compounds such as amino acids and amines; different NHt cells have different second-best compounds (Johnson et al., 1984). We tested the tuning spectrum of 15 NH, receptors using Asp, Arg, Hyp, Glu, Bet, TMO and TMA at a concentration of 0.3 mM. Of these compounds, only Bet, Glu and, to a lesser extent Hyp, could stimulate NH( cells. On average, these compounds elicit 1096 or less of the response to NR, at equimolar concentration. In cross-adaptation experiments we searched for a NH4 cell, determined whether Bet, Glu (or Hyp) was its second-best compound, and then used either Bet or Glu as adapting compound. Subsequently, we recorded two NH4 stimulus response functions (0.003 - 3 mM NJ^Cl): one in artificial seawater (ASW), and one in a background of ASW plus various concentrations of the second-best compound (0.003-3 mM Bet or Glu). When no second-best compound was found, either of the two compounds was used in the background. This protocol allowed us to study the effects of cross-adaptation on receptors with and without these second-best compounds. This research was supported by the Whitehall Foundation and NSF (BNS-8512585).

Single channel recordings from chemosensory neurons on the crayfish walking leg
Hanns Hart Physiologisches Institut, Technischen Universitdt MOnchen, 8000 MOnchen, FRG Single channel currents were recorded from chemosensory neurons by means of a GG-seal patch clamp in cell-attached and cell-free modes. Channels were activated by nicotinamide and analogous substances in the pipette. Single channel current was about 3 pA at resting potential; it changed linearly with potential shifts snowing a conductance of 2 5 - 3 0 pS and a reversal potential of +15 mV. AT 5 x 10"' mol/1 nicotinamide, the mean open time was 1 2 ms, while at 10~* mol/1 bursting activity was elicited. In outside-out patches, doseresponse curves were determined which were similar to those obtained from

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Nicotinamide -80mV 5x10" 6 mol/l

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action potential frequency of single afferent fibres [Hart and Schmiedel-Jakob (1984) J. Comp. Physioi, 154, 855]; also the sequence of effectivity of drugs was the same. The results indicate the presence of chemosensory receptor molecules also in the membrane of the cell body. Supported by the DFG.

Processing of olfactory information in the Colorado potato beetle


R.de Jong and J H.Visser Department of Entomology, Agricultural University, PO Box 8031, 6700 EH Wageningen, The Netherlands Adult Colorado potato beetles, Lcptinotarsa decemlineata, can distinguish their food plant, Solatium tuberosum, from others by perceiving differences in plant odour compositions. Some general green leaf volatiles, like cw-3-hexen-l-ol, c/.r-3-hexenyl acetate, iranj-2-hexen-l-ol, jra/u-2-hexenal and 1-hexanol, are thought to play an essential role in plant odour recognition. The beetle's receptor neurones showed overlapping reaction spectra when stimulated with some of these leaf odour components. Response characteristics of olfactory neurones in the antennal lobe can be divided roughly into two groups. One group (A) containing neurones which are not very specific for the tested leaf odour components, and another group (B) with neurones which show specific responses to one of these components. When the antennal receptors were stimulated by a potato leaf extract, group A neurones of the antennal lobe responded, while group B neurones hardly changed their spontaneous firing activities. It is concluded that the two groups of neurones differed in two respects: (i) their specificities of responses to individual components, and (ii) the presence or absence of response to a potato leaf extract. Since the potato leaf extract can be considered as a mixture of odour components, the lack of response in group B neurones to such a mixture implies an important role of inhibition as a mixture effect. R.de Jong was supported by the Foundation for Fundamental Biological Research (BION), which is subsidized by the Netherlands Organization for the Advancement of Pure Research (ZWO).

Temporal coding of alkali chlorides in blowfly salt receptor responses


F.W.Maes and G.Harms Department of Animal Physiology, State University Groningen, PO Box 14, 9750 AA Haren, The Netherlands Blowflies Calliphora vicina are able to discriminate between some alkali chlorides in a classical conditioning paradigm [Maes and Bijpost (1979) J. Comp. Physioi., 133, 53]. The information about salt taste quality, necessary for this behaviour, may be contained in an ensemble code [Maes (1980) Olfaction and Taste VII, p. 123; Maes and Ruifrok (1986) J. Comp. Physioi. A, in press]. The present contribution considers the alternative possibility of temporal coding, through differences in the time course of adaptation of the phasictonic salt receptor response. [A more detailed account is given in Maes and Harms (1986) J. Comp. Physioi. A, in press.] Salt cell responses of 89 labellar taste hairs were obtained to a reference stimulus (1 M KC1) and a doubling concentration series of one of the alkali chlorides (LiCl, NaCl, KC1, RbCl or CsCl). Times of spike occurrences were measured with an 'electronic ruler' at 0.2 ms resolution, for the first 500 ms of response. Responses were then converted into 'adaptation curves' of momentary firing frequency versus time. Mean

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ECRO VH: Abstracts adaptation curves for the 1 M KC1 reference fitted neatly into the array of curves for KC1, RbCl and CsCl, but less so for LiCl and NaCl. Quantification of adaptation curve shape, necessary to assess the significance of the latter deviations, was obtained by using two parameters: the shape factor SF (ratio of spike rates in the 0125 msand 125-500 ms intervals) and the mean spike rate SR over the 0 - 5 0 0 ms interval. The responses to the concentration series of an alkali chloride yielded a trajectory in the SFSR response plane. The point representing the 1 M KC1 reference appeared to be on the trajectories for KC1, RbCl and CsCl, but significantly outside those for LiCl and NaCl. These deviations are presumably due to differences in diffusion velocity of salts through the terminal pore into the 'tip chamber' of the hair. Whether the observed shape differences are used in a temporal coding scheme for salt taste quality is not very probable, as the differences are small and the decoder would require a rather complex neural network.

Comparative electrophysiological studies on olfaction in tsetse flies (Glossina spp.)


C.J.Den Otter and T.Tchicaya1 Department of Animal Physiology, Postbus 14, 9750 AA Haren, The Netherlands, and iIEMVT, 10 rue Pierre Curie, 94704 Maisons-Alfort Cedex, France Vale [Bull. Entomol. Res., 70, 563-570 (1980)] found that odour-baited traps are valuable aids for sampling populations of tsetse flies and offer promise as a means of tsetse control. However, behavioural and field trapping experiments are very time-consuming. Electrophysiology offers a relatively rapid method of screening odours for their activity at the receptor level. We made comparative studies on the responses of olfactory receptors (EAGs) in different species of tsetse flies to substances which were found to be attractive or repellent to Glossina m.morsitans and G. pallidipes (Vale, 1980). Species tested included both males and females of G. m.morsitans, G. austeni, G. palpalis gambiensis, G. tachinoides and G. f.fusdpes. Substances applied were l-octen-3-ol (highly attractive), acetone (highly attractive), 4-heptanone (attractive), 3-nonanone (repellent), each at five different concentrations. In addition, behavioural studies have been initiated in the laboratory to establish the relative attractive or repellent effects of odours. Results of both electrophysiological and behavioural experiments are presented.

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Host and non-host plant chemicals influencing the oviposition behaviour of several herbivorous insects
R.SchSni, E.StSdler, J. J.A.Ren wick1 and C.Radke1 Eidg. Forschungsanstalt, CH-8820 Wadenswil, Switzerland, and lBoyce Thompson Institute at Cornell University, Ithaca, NY 14853, USA The influence of chemicals extracted from cabbage leaves on the oviposition behaviour of the cabbage pest insects Delia radicum, Trichoplusia ni, Pieris rapae, Pieris brassicae and Mamestra brassicae and the carrot fly Psila rosae was tested in specific laboratory bioassays using natural and artificial egg laying substrates. Active raw extracts stimulating or deterring oviposition were separated with analytical techniques into purified fractions. Attempts were made to identify the active chemical stimuli contained in the isolated fractions. Purified extracts were also used to determine sepcific receptor cells responsible for the perception of the active components. A comparison of the collected data shows that the decision to accept or reject a plant as a suitable host is not uniquely based on some few key stimuli, e.g. the glucosinolates, but rather on a large variety of stimulatory and inhibitory plant chemicals acting together. In addition the active host plant 'blend' seems to be specific for each species. This paper will be published in extenso in the Proceedings of the 6th International Syposium on Insect-Plant Relationships, held in July 1986 at Pau, France.

Electrophysiological recordings used as bioassay for the isolation and identification of a non-volatile pheromone
Erich Stadler, Reto Schoni, Jakob Hurter and Ernst Boiler Eidg. Forschungsanstalt, CH-8820 Wadenswil, Switzerland The water-soluble oviposition-deterring pheromone (ODP) is deposited by the female cherry fruit fly Rhagoletis cerasi on the cherries after oviposition. The ODP was found to be present in the faeces of mature females

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ECRO VII: Abstracts and was isolated and identified by Hurter et al. (1). The biological activity of extracts and fractions was determined by comparing the nerve impulse activity of the ODP best cell in the tarsal D-sensilla of the fly. The results of this electrophysiological bioassay were verified with behavioural observations and proved to be reproducible and reliable. The suggested structure of the pheromone is N(1503-glucopyranosyl)-oxy-8hydroxypalmitoyl)-taurine. (1) Hurter.J., Boller.E.F., Stfidler,E., Blattmann.B., Buser.H.R., Bosshard.N.U., Damm.L., Koslowski, M.W., Schdni.R., Raschdorf.F., Dahinden.R., Schhimpf.E., Fritz.H., Richter.W.J. and SchreiberJ. (1986) Oviposition deterring pheromone in Rhagoleris cerasi L.: purification and identification of the chemical constitution. Experientia, 42, in press.

Behavior-stereotypes of feeding and those displayed in rejection of aversive-tasting food by the freshwater prawn: Macrobrachium rosenbergii
Jacob E.Steiner and Sheenan Harpaz1 Department of Oral Biology, Hadassah Faculty of Dental Medicine, ^Department of Zoology, Life Sciences Institution, The Hebrew University of Jerusalem, Israel Previous investigations carried out on vertebrates in general, and on mammals in particular, showed that fixed behavior patterns reliably indicate acceptance and aversion in response to chemical stimuli. The present study focuses on an invertebrate, a decapod crustacean. Food search and food intake behavior stereotypes of the fresh-water prawn Macrobrachium rosenbergii were studied on a sample of 27 adult animals of both sexes in the intermolt phase of the molt cycle. Fooddeprived animals were stimulated with an actual food item, to which they were accustomed, or with small samples of different concentrations of the chemoattractant betaine-HCl, or with food pellets adulterated with quinine-HCl. The behavioral displays induced by all these stimulants were videotaped and recordings were evaluated by two independent viewers. Behavior analysis was based on quantitative assessment of antennular flicks; on that of probing movements by pereiopods as well as pereiopod lifting movements to the labial region. Results revealed that the food search and food intake behavior sequences induced by actual food can be evoked in an identical manner when only betaine simulated the nutrient. The betaine-induced features show a clear dependence on stimulus intensity. The quinine-adulterated food sample triggered all the initial sequences of food search and food lifting behaviors apparently due to stimulation chemoreceptors, located on antennae and antennulae. Detection of the aversive taste most probably by labial chemoreceptors induced a different set of motion features, resulting in food rejection (dropping or active expulsion) accompanied by typical oral-cleaning movements.

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Single cell responses combined with gas chromatographical separation of host volatiles in bark beetles
B. A .T0mmeras, H.Mustaparta, O.Gederaas1 and T.Anthonsen1 Department of Zoology and ^Department of Chemistry, University of Trondheim, Norway Bark beetles are dependent on odours of host trees to find breeding materials. Previous electrophysiological studies of single olfactory receptor cells have shown that these insects possess a large number of cells which are strongly activated by vapour of host tree bark and do not respond to other biological signals, e.g. pheromones (1). The aim of this study was to determine which constituents of the host volatiles are the effective stimulants for the receptor cells in bark beetle species. Single cell recordings were combined with gas chromatography of host odour extracts. The column of the gas chromatograph (g.c.) was split at the end, leading one half of the effluent to the g.c. detector, and the other half over the insect antenna (2). Screening for sensitivity to 'whole' bark vapour, whole extract vapour (of various bark materials made by different methods) and separated g.c. fractions, revealed three patterns of responses: (i) excitation by a large number of cells to whole bark vapour, but not to any of the extracts made; (ii) excitation to bark vapour, to one or more extracts and to specific fractions of the same extract after g.c. separation; (iii) excitation by a few cells to bark vapour and to one extract, but not any single fraction of it. The cells responding to single fractions (ii), responded generally to minor constituents. Responses by a few cells to main constituents, such as pinenes, were very weak.

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ECRO VII: Abstracts The results indicate that host material quality as well as extraction methods are important for obtaining effective olfactory stimulants. Furthermore, it appears that the effective compounds are among the minor constituents of host volatiles, and that the receptor cells may possess a high degree of specialization for such compounds. The possibility that synergism between host odour constituents also may take place on the receptor cells should, however, not be excluded. 1. Mustaparta.H. (1979) In Ritter.F.J. (6d.), Chemical Ecology: Odour Communication in Animals. Elsevier/N. Holland. 2. Wadhams.L. (1982) Z.Naturforsch., 37, 947-952. The study is financed by the Norwegian Research Council of Technology, NTNF, and Borregaard Industries Ltd., Norway.

Importance of signal-to-noise ratio and absolute stimulus concentration for cumulative self-adaptation in ehemoreceptor cells
Rainer Voigt, Carl L.Merrill and Jelle Atema Boston University Marine Program, Marine Biological Laboratory, Woods Hole, MA 02543, USA The lateral antennular flagellum of the American lobster functions as an olfactory organ, and is of critical importance for orientation in odor plumes. The patchy nature of odor plumes makes it likely that this organ and its receptor cells detect pulsed stimuli in a chemically noisy background. A distinct population of these receptor cells is narrowly tuned to taurine. Cell sensitivity varies due to (i) self-adaptation to a constant background; (ii) cumulative self-adaptation to repeated stimulus pulses; and (iii) cross-adaptation due to mixture suppression. We have started to characterize the time courses of adaptation and disadaptation of receptor cells using series of standard 1 s stimulus pulses with varying signal-to-noise ratios. The temporal stimulus pulse profile was determined by measuring the change of conductivity of flowing deionized water (20 ml/min) after injection of 100 /A of 1 M NaCl solution over 300 ms. The stimulus chamber allowed 5 s interpulse intervals without interference of the previous stimulus. Single cells were identified electrophysiologically with 10~* M taurine. A series of five pulses was applied in 10 s intervals for one of several concentrations (10~ 3 10~'M) in different backgrounds (10~410~7 M). Combinations of stimulus and background concentrations provide similar signal-to-noise ratios at different absolute concentrations and vice versa. Preliminary results suggest that at different background levels the same signal-to-noise ratio resulted in similar responses, including similar cumulative adaptation. Greater signal-to-noise ratios caused stronger responses and showed greater cumulative adaptation while smaller ratios caused weaker responses and less adaptation. Thus, signal-to-noise ratios and no absolute stimulus levels were predictive of the responses of receptor cells in different backgrounds. Supported by NSF grant BNS 8512585.

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VI. Poster Presentations 1. Olfaction A comparison of odor perception in smokers, non-smokers and passive smokers
R.AhlstrSm, B.Berglund, U.Berglund1, T.Engen2, and T.Lindvall3 University of Stockholm, 'Royal Institute of Technology, Sweden, 1Brown University, New York, USA, ^National Institute of Environmental Medicine, Stockholm, Sweden The olfactory perception of matched groups of 26 smokers, 26 non-smokers and 15 passive smokers was examined psychophysically with two substances, n-butanol and pyridine. Different psychophysical functions were obtained from these groups with the method of magnitude estimation. The smokers and passive smokers perceived all concentrations of n-butanol to be weaker than did the non-smokers. In the case of pyridine, to which the smokers are more exposed from tobacco smoke, there is an indication of a perceptual deficit similar to loudness recruitment. There were no appreciable differences between the smokers and non-smokers

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ECRO VD: Abstracts for high concentrations, the sensitivity was less and the psychophysical functions steeper for smokers than non-smokers for relatively low concentrations. The results are evaluated in terms of information about the subjects' response criterion, and the explanatory possibility of sensory deficit versus habituation are considered.

Frog E.O.G. in low conducting solutions


I.Andriason and J.Leveteau Laboratoire de Neurophysiologie comparie, 4 Place Jussieu, 75230 Paris, France Previous experiments had demonstrated that negative E.O.G. amplitude increases when olfactory mucosa is perfused with a low conducting solution. This strongly favoured the idea of a current generator in series with the mucus compartment. This inward current generator should be located above the epithelial surface in order to account for the known magnitude of the E.O.G.-related electrical events. When the perfusing solution contains sucrose (224 mM), KHCO, (2 mM) and NaCl (2 mM), the E.O.G. elicited by 1 mM isoamylacetate was ~ 7 mV in amplitude. Replacing 2 mM NaCl with 1 mM CaClj resulted in a 64% increase of E.O.G. amplitude in spite of the fact that the solution conductivity was almost unchanged. Replacing Na ions in the other solutions, without appreciably modifying resistivities induced the following changes: Ions Concentrations (mM) 1 0 - 4 S/cm Relative E.O.G. amplitudes Ca 1 4.4 1.6 Ba 1 4.6 1.4 2 4.5 1 3.8 0.9 Na Co 1 Cd 1 4.3 0.3

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Addition of 0.1 mM EGTA in Ca-free solution resulted in relative E.O.G. amplitude of 0.5. All the observed effects were reversible except for cadmium solution. These observations suggested that Ca ions were charge carriers as were Ba, Na and Co ions. In addition Ca ions potentiated the E.O.G.

Behavioral and neural development of ferrets raised under geraniol overexposure


Raimund Apfelbach UniversitOt Tubingen, Institut far Biologie III, Aufder Morgenstelle 28, 7400 Tubingen, FRG In the carnivorous ferret (Mustela putoris f.furo) searching for prey can be reliably elicited by known food odors, unknown odors are not reacted to. The reactions to even two familiar odors are not necessarily the same; preference for a specific prey odor is due less to the length of prior feeding experience than to exposure during a sensitive phase during post-natal ontogeny, occurring between day 60 and 90. In order to study the influence of early olfactory experience on the behavior and on the developmental changes in the olfactory bulb (OB), the effects of early olfactory deprivation on these parameters were investigated. Olfactory deprivation was induced by rearing litters in an artificial environment saturated with geraniol odor. The behavioral data indicate a preference for geraniol odor in the animals raised under geraniol overexposure, while control animals of the same age showed aversive behavior when presented with this substance. The readiness of the olfactory-deprived animals to respond towards novel odors seemed also to be inhibited; the underlying causes for this behavioral change are not yet known. The developmental stage of the OB was assessed by quantifying the number of spines on the external dendrites of granule cells (Golgi staining method). The results reveal that the relative state of odor deprivation caused by geraniol odor overexposure does not impart its effects on the initial phase of growth where spine number is increasing, but rather on the latter phase of growth where this parameter is decreasing. This indicates not only that this later phase of granule cell development in the OB is a plastic and vulnerable phase, but that early olfactory experience may regulate the normal decline in this parameter. Further studies are needed to investigate the underlying causes of the overshoot and decline in neural structures and any relationship that may exist between the latter and behavioral development. Supported by the Deutsche Forschungsgemeinschaft (Ap 14/8-6).

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ECRO VII: Abstracts

Control of male rat's olfactory bulb reactivity to sexual odours


Jean-Marc Azerad and Rimi Gervais Laboratoire de Physiologic neurosensonelle (UA CNRS 180), University Claude-Bernard, F-69622 Villeurbanne Cedex, France Several electrophysiological studies have provided evidence that, in the rat, the olfactory bulb responsiveness to some odours is related to their biological significance. This effect was shown to involve the action of fibers of central origin. Behavioural studies have demonstrated that sexually experienced male rats prefer urine odor from receptive females (RU) rather than that from non-receptive females (N RU). This preference disappears after castration. We attempted to test the hypothesis that this behavioural preference is associated with a different bulbar reactivity towards these two odours. We developed a new type of multiunitary recording from mitral cells combined with a spectra] analysis of the complex signals. In unanaesthetized animals responses to odour stimulations clearly appeared in the 80 Hz frequency band, irrespective of the coordinates of the electrode implantation sites in the mitral cell layer. This activity was elicited significantly more often by RU than by N RU. The difference in reactivity towards these urine odours disappeared after section of the olfactory peduncle and after castration. The involvement of the actions of centrifugal fibers (to be identified) and of steroid hormones may therefore be assumed.

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Optical mapping of the piriform cortex electrical responses to lateral olfactory tract stimulations in the rat
M.Cattarelli and L.B.Cohen1 Physiologie Seuro-Sensorielle, University Claude Bernard, 69622 Villewbanne Cedex, France, and ^Department of Physiology, Yale University School of Medicine, New Haven, CT 06510, USA It has been shown that optical recording methods can be used for monitoring electrical activity in mammalian cortices (1). We have used this method for studying the spatial distribution of the activity induced in the piriform cortext by electrical stimulations of the lateral olfactory tract (LOT) of the rat. Surgical procedures including tracheotomy, jugular vein cannulation and exposure of the latero-basal brain surface were performed on Wistar SPF male rats anaesthetized with equithesine. The anaesthesia was continued during the recording. In addition the animals were curarized and artificially ventilated. Piriform cortex was stained with a voltage-sensitive dye, either RH414 or RH795 kindly supplied by R.Hildcsheim and Dr A.Grinvald. Cortical fluorescence was measured with a 124 element photodiode array using epi-illumination while electrical stimulations were delivered to the LOT. Eight or 16 trials were usually averaged. A five-wire electrode located on the anterior LOT allowed us to deliver various spatially localized stimulations to this nervous pathway. Optical signals with a duration at half height of from 15 to 100 ms were recorded in response to LOT stimulations. In some preparations their shape and distribution varied according to the location of the LOT-stimulating electrodes. This result suggests a somewhat organized spatial projection of the LOT on the piriform cortex. 1. Orbach.H.S., Cohen.L.B. and Grinvald.A. (1985) J. Neurosd., 5, 1886-1895. Supported by PHS Grant number NS08437, Centre National de la Recherche Scientifique, NATO and Fondation pour la Recherche Medicale.

Response properties of olfactory bulb neurons to odour concentrations in awake freely-breathing animals
Michel A.Chaput Laboratoire d 'Electrophysiologie el Laboratoire Associt au CNRS de Physiologie Neurosensorielle, Universitt Claude-Bernard, Lyon 1, 69622 Villeurbanne Cedex, France Single-unit extracellular activity was recorded in or near the mitral cell layer in the olfactory bulbs of awake rabbits. Five odorants were used as stimuli: acetophenone, cineole, isoamyl acetate, p-cymcne and cyclohexanone. They were delivered at 1 0 ~ \ 3 x 10"*, 10" 1 and 10~2 of the saturated vapour pressure at 21C. They were presented in increasing concentration order. Stimulations lasted 10 s and were separated by at least 90 s.

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ECHO VII: Abstracts Responses were recorded from 65 presumed mitral cells. They were analyzed by processing separately the inspiration-related (INSP-) and the expiration-related (EXP-) activity since we have previously shown that olfactory bulb neurons respond to odorants by a respiratory phase-related reorganization of their discharge. They increase their activity during inspirations and decrease it during expirations. Responses were characterized by their magnitude during each phase, i.e. by the mean change in INSP- and in EXP-firing frequency with respect to the activity recorded during the corresponding phases before each stimulus delivery. They were also characterized by their type: excitation, inhibition or null response. The analysis of response types shows that increasing odor intensity significantly increased the olfactory bulb responsiveness. Twenty-five per cent of the stimulations evoked a response for 10~4; 34% for 3 x 10" 4 ; and 69% for 10~2. This enhanced responsiveness resulted from an increase in the number of responses characterized by an INSP-actvation only, by an EXP-inhibition only; or characterized by an INSPactivation associated with an EXP-inhibition. Response magnitudes were used to compare the response profile of the set of neurons across concentrations. Profiles evoked by an odorant were generally similar for 10~\ 3 x 10~* and sometimes 10"', and were different for 10~ l . This last result raises the question of the independence between the quality and the intensity coding in the olfactory bulb of awake animals.

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The effects of post-partum olfactory stimulation on further food preference in the female albino rat preliminary results
Michele Cormareche-Leydier, Arlette Duveau and Jacques Chanel Laboratoire de Physiologic neurosensorielle, CNRS, University Claude BernardJLyon, F-69622 VMeurbanne Cedex, France The effects of post-partum olfactory stimulation on food preference were investigated in four groups of four female rats. Olfactory stimulations were provided by daily painting abdomen skin with an isoamyl acetate alcohol solution (50% isoamyl acetate + 50% ethanol) during six successive days. In group 1 painting was applied from day 1 to day 6 following parturition, in group 2 from day 7 to day 12 and in group 3 from day 13 to day 18. A fourth group of four females, painted with the solvent only (two of them from day 1 - 6 and the other two from day 7 - 1 2 following parturition) were used as controls. Thirty to 60 days later, the females were given the choice between usual food (UAR 180) and usual food flavored with isoamyl acetate, both provided ad libitum in individual cages for seven successive days. Flavored food ingestion was significantly increased in group 1 rats. Group 2 ate similar amounts ofilavored and unflavored food. Group 3 behaved as did the controls and preferred unflavored food. The mechansims according to which early post-partum olfactory stimulation could induce changes in food preference in the rat are under investigation.

Actions of the efferent innervation on goldfish olfactory bulb units during odour stimulation
T.Fischer, H.P. Zippel and D.Schild Physiologisches Institut der Universitat, Humboldtallee 23, D-3400 Gdttingen, FRG The effects of the efferent pathways on the inner bulbar information processing have frequently been studied by electrically stimulating the olfactory tract and other structures participating in the efferent innervation of the olfactory bulb. From these investigations it is evident that the centrifugal fibre systems act inhibitorily on the mitral cell activity via the granule cells. Virtually nothing is known about the role of the efferent pathways on olfactory bulb units during repetitive natural stimulation of the olfactory mucosa. In the present investigations recordings were made from mitral cells during odour application before and during blockage of the efferent fibre systems by cooling the olfactory tract down to 1 C. The same odour has been presented for at least 30 s, 40 times before and then 40 times during cooling. Each stimulus period was preceded by a stimulus-free interval (at least 30 s), during which a steady stream of tap water was applied. This experimental procedure allows the investigation of activity changes by statistical means. The results can be summarized as follows, (i) In the mean overall experiments, cooling had no general effect on static activities (long term activity: 12 - 3 0 s) in the stimulus-free and the stimulus interval: in comparison with the static activity recorded before cooling, the static activity was higher (excitatory), unchanged (indifferent) or lower (inhibitory). From this finding changes in mitral cell static activity during efferent

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ECRO VII: Abstracts blocking are obvious, but no shift in the expected direction (i.e. a general disinhibition of the mitral cells) has been observed, (ii) Blocking the efferent fibre systems has, in contrast, a significant (Wilcoxon-ranktcst, P = 0.025) influence on the dynamic response ( 0 - 6 s after stimulus onset): in comparison with the effects recorded before cooling, the dynamic response is reduced during the efferent blockage. This effect is evident in both transient responses, from water to stimulus phase and vice versa. The recorded effects could be interpreted as follows: blocking the efferent fibre systems to the olfactory bulb influences the information processing predominantly during changes in the olfactory environment, i.e. one effect of the efferent pathways under biological conditions is an enhancement of the animals' ability to detect odour sources different from the permanent olfactory environment.

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Projections of the fila olfactoria in catfish, Ictalurus nebulosus


T.Lago-Schaaf, Y.F.Tsai and P.Bruckmoser Zoologisches Institut der UniversitOt, Scidlstrassc 25, 8000 MOnchen 2, FRG Parallel to electrophysiological studies in the olfactory bulb (OB) the projections of the olfactory mucosa (OM) to the OB in catfish were investigated. The OM was subdivided into four quadrants and HRP was applied to the central lamellae of each quadrant. The receptors of both lateral quadrants send their axons through the lateral ON GON), the receptors of both medial quadrants through the medial ON (mON). In contrast to the salamander (Dubois-Douphin.M. et al., 1981) the fibers coming from a discrete area of the OM do not remain together on their way to the OB; rather they distribute over the whole surface of the ON. Just before entering the OB the ION and the mON cross partially to the opposite sides. The projections of the four quadrants are identical with our method. There is no topographical relationship between the OM and the OB with the exception of a group of 2 0 - 3 0 secondary neurons in the caudal OB (Tglb) receiving projections only from the rostral quadrants. Catfish have a good sense of smell and their complex social life is regulated by olfaction (Atema,J. et al., 1969). Thus the diffuse olfactory projections are as unexpected as the finding that the OB has no more than 200300 secondary neurons. Other macrosmates have many more secondary neurons and distinct somatotopy in this area. The role of the Tglb in sexual behavior and the possible meaning of the diffuse projection are discussed.

Multi-site electrical stimulation of the olfactory bulb in the rat: discriminative and perceptive properties
Anne-Marie Mouly and Andr6 Holley Laboratore de Physiologic neurosensorielle (UA CNRS 180), F-69622 Villeurbanne Cedex, France Low-intensity electrical stimulations of the olfactory bulb were delivered through arrays of electrodes located in various sites of the mitral cell layer in unrestrained rats. This experimental model aims at simulating the main features of the olfactory message by creating spatially-distributed configurations of neural activity. In a two-choice paradigm, a combination of stimulation sites was associated with a palatable solution delivered to water-deprived rats while another combination was associated with a non-palatable solution. These multisite stimulations proved to be sufficient cues for guiding the animals' choice even when electrode patterns to be distinguished overlapped each other to a large extent. Perceptive effects induced by stimulating electrodes separated by no more than few microns were still discriminated. The animals' perception of multi-site stimulation patterns seems to be analytical rather than synthetic: the rats could recognize a given stimulation site when this site was stimulated together with several others. Individual characteristics of bulbar stimulation appear to be perceived absolutely rather than differentially: identification of a multi-site pattern was still possible when this pattern was presented alone in test sessions, rather than concurrently with another one. A good retention of the discrimination learning of specific stimulation patterns was observed: animals could identify stimulation patterns after complete interruption of the training for 17 days.

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ECRO VH: Abstracts

Pheromonally induced intermale aggression in laboratory mice: genetic aspects and the model's prospects
S.N.Novikov, N.A.Djuzhikova, M.B.Pavlova and V.V.Babalyan Pavlov Institute of Physiology, USSR Academy of Sciences, Leningrad 199164, USSR There are two main ways for the study of the genetic basis of pheromonally mediated aggression in male laboratory mice. The first one is to evaluate the genetic mechanisms controlling biogenesis of these substances. The second way involves genetic study of pheromonal actions on the organism. Assuming that volatile steroids play an important role in intermale agonistic behaviour (e.g. Ingersoll et al., 1982) we present some evidence for genotype-specific j3-glucuronidase (EC 3.2.1.31) mode of action on the immobilized (conjugated) propheromone in mice (Novikov and Djuzhikova, 1986). We supposed that strain differences in the spectrum of excreted and deconjugated steroid glucuronides with pheromonal potency were defined by the activity of 3j3-hydroxystcroid dehydrogenase (EC 1.1.1.145) and pH value. The presented genetic regulation scheme of aggression promoter activity is based on the action of the Mup complex (chromosome 4), Gus complex (chromosome 5) and 3/3-hydroxysteroid dehydrogenase activity. It seems likely that the most fruitful method for the study of genetic control of an organism's reactivity to pheromones is analysis of genetic control of catecholamine biosynthesis in different limbic structures. The proposed model is the first attempt to evaluate the links between localized genes and aggression in Mus musculus.

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Quantitative structure activity relationship in olfaction


Francois Patte and Paul Laffort Laboratoire de Physiologie de la Chimiortception CNRS, F-91190 Gif-sur-Yvette, France The quantitative relationships we have found between olfactory activity and molecular structure have not really been improved for several years (ECRO HI, 1978). They are similar to those obtained by some colleagues with other approaches: significant, but not sufficiently high to explain the entire phenomenon. For this reason, we have tried to improve, during these last years, the accuracy of the three fields needed for correlations, i.e. data banks in molecular structure and biological activity, as well as a better definition in models of olfactory activity. The work in progress concerns on one hand another characterization of molecular structure, based on a topological approach, and on the other hand, the determination of confusion matrices from the proboscical reflex in the honey-bee. The results concern the quantitative interaction model in olfactory mixtures, the demonstration of olfactory perception for krypton and methane in hyperbaric atmosphere, the EAG doseresponse curves in the honey-bee for 59 substances and the interaction of quantity and quality of the stimulus in proboscical reflex.

Ultrastructural features of internal granule cells in the rat olfactory bulb


C.Reyher Institute of Neuroanatomy, Freie UnivtrsitOt, Berlin, FRG The internal granule cells of the main olfactory bulb are thought to be one of the inhibitory elements which modulate signal processing of the olfactory bulb neurons. Thus, the granule cells contact the secondary dendrites of the output neurons, mitral and tufted cells, via reciprocal synaptic membrane complexes. These synapses occur at the characteristic of these neurons, the gemmuli, which arc also identified as the postsynaptic targets for the incoming fibers arising in higher olfactory and limbic brain structures, i.e. centrifugal afferents. In order to define the apposition between these neurons we used the tannic acid fixation technique and analyzed sections prepared for the electron microscope. In this material the granule cells situated deep in the mitral cell layer occur as neurons with a size ranging from 7 to 11 fun in diameter. Most of them had a spherical or fusiform shape and relatively large nucleus surrounded by a small cytosplasmic rim. The nuclei appearedrelativelydark because of the condensed chromatin material at the peripheries and due to the presence of prominent nucleoli. Gap junctions occur between neighboring perikarya. The ultrastructural organization of these presumptively GABA-ergic interneurons indicates that there may

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ECRO VII: Abstracts be cell-cell communciation between the neighboring perikarya. Since granula cell inhibitory synapses are thought to mediate lateral inhibition of afferents throughout the olfactory bulb, the presence of gap junctions between neighboring granule cells may function to synchronize activity among subpopulations of these neurons and serve to coordinate the timing of lateral inhibitory effects. In addition, gap junctions may reflect the relative immaturity of granule cells which are still being generated well into adulthood, since gap junctions are a characteristic of immature cells.

Brief olfactory stimulations in humans


Catherine Rouby, Michel Vigouroux and Andrd Holley Laboratoire de Physiologie neurosensorielle (UA CNRS 180), University Claude-Bernard, F-69622 Villeurbanne Cedex, France We present details of a program to study the perceptual properties of brief olfactory stimulations in man. The whole progTam includes the use of two identical and separately adjustable stimulators, allowing the application of short stimulations (range 15-750 ms) with a varying delay. A stimulator has been realized and used to investigate the relationships between duration of stimulus and perceived intensity, for different concentration levels, (i) The stimulation is triggered by the subject's sniff, (ii) The weak flow-rate (200300 ml/nun) of odorized air makes it impossible to detect the stimulation by the somaesthesic way. (iii) The odor is injected at high speed in the nostril: this allows the odorized air puff to reach the olfactory mucosa, even with the shortest durations, (iv) By measuring the inhalation flow-rate, we can assess the dilution of the stimulus in the sniffing air flow, (v) By recording the position of the stimulation within the sniffing episode, we can assume its efficiency on perceived intensity.

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Formation of inclusion complexes of starch with flavor compounds


M.A.Rutschmann and J.Solms Department of Food Science, Swiss Federal Institute of Technology, Zurich, Switzerland The complex formation of starch with the flavor compounds methone and decanal was studied in a ternary system. The reaction was followed by iodine binding determinations and by quantitative analysis of the complex composition. The inclusion process is a slow one and approaches a final level where all active sites of the amylose molecule are occupied. The degree of complex formation of the different ligands in mixed systems varies with concentrations and shows interesting effects with apparent synergistic and antagonistic effects. These variations are certainly due to the specific characteristics of the ligands. However, changes in the conformation of the amylose helix with an adaption to the dimensions of the ligands also seem to be of great importance.

Reproducibility of mitral cell responses in goldfish


D.Schild and T.Fischer Physiologisches Institut der University, Humboldtallee 23, D-3400 Gottingen, FRG Implicitly, it has often been assumed that mitral cell responses of the olfactory bulb are reproducible; only one or a few runs were therefore taken in order to describe a stimulus response. However, it has recently been shown that the dynamic component of EOG responses changes considerably during some stimulus applications (van Aset al., 1985); further, Gray et al. (1984) provided evidence for plastic processes within the olfactory bulb. It thus seemed worthwhile to study the reproducibility of extracellular recordings from mitral cells. Using the olfactometer described previously it was possible to record 40 stimulus responses to the same stimulus, whereby stimulus duration as well as interstimulus duration was 30 s. During the interstimulus periods water was delivered to the mucosa. Tubifex extract (10"* M) and amylacetate (10~*) were used as stimuli. It has been reported qualitatively [Schild and Zippel (1986) /. Comp. Physiol. A, in press] that the responses are sometimes not stable in the course of the 40 experimental repetitions (runs). These response changes across the runs might indicate synaptic changes within the bulb or the effect of efferent control on the bulb. In this study we analyse the activity changes across the runs statistically: the regression between activity per bin group and run index was calculated, whereby bins of 1 s length were chosen. By this method one can detect activity changes in the course of the runs without assuming any pattern features.

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ECRO VD: Abstracts The assumption of a dynamic and a static response component as pattern features (Schild, ECRO VII, 1986) allows a further method of checking the reproducibility of recordings: four consecutive runs of a recording are respectively considered as one ensemble; then the dynamic and static response activity are compared to the pre-stimulus activity. In this way, the dynamic and static response can be observed in runs 1 - 4 , 5 - 8 , 9 11,..., 36 - 4 0 . The activity changes of both the dynamic and static response tend to zero after a certain number of runs; two types of changes occur: in some cases a significant response observed at the beginning of an experiment vanishes after some runs (the system adapts to 'no reaction'); in other cases, no response is observed at the beginning of an experiment, but a response gradually evolves in the course of the runs (the system adapts to 'optimal reaction').

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Glomerular activation patterns elicited in the olfactory bulbs of AKR and C57 Bl/6 mice by odorant stimulations
It has been reported that C57 Bl/6 mice show a higher detection threshold to iso-valeric acid than do AKR mice (Wysocki et al., 1977). We have studied the glomerular activation patterns elicited by odorant stimulation in C57 Bl/6 and AKR mice using the ["C]2-deoxyglucose labeling method. Free moving animals were exposed during 45 min to an alternatively odorized-deodorized air environment. In the first group of experiments, the tested stimuli, amyl acetate or iso-valeric acid, were delivered at 10~* M concentration levels. The autoradiograms obtained from serial sections of the olfactory bulbs were submitted to densitometnc analysis and the glomerular activation maps were reconstructed according to the method developed by Royet etal (1986). The comparison of the glomerular activation patterns demonstrates that: (i) the spatial organization of the activation patterns depends clearly on the odorant quality of the stimulus; (ii) the characteristic patterns elicited by one odorant were found to be similar in the two strains; (iii) in the two strains, numerous neuroreceptor cells which are activated by iso-valeric acid stimulations converge on a restricted area of the glomerular layer. Further experiments, using various odorant concentrations, were pursued and the correlative variations of the glomerular activation patterns were observed. The results are discussed, considering the origin of the specific anosmia to iso-valeric acid in C57 Bl/6 mice. G.Sicard, J.P.Royet and F.Jourdan Laboratoire de Physiologic neurosensorielle, Vniversiti Claude-Bernard, F-69622, ViUeurbame Cedex, France

Effects of ferulic acid on the aroma quality of orange juice during storage
Benjamin J.Striem, Michael Nairn, Joseph Kanner1 and Hanna Peleg Department of Biochemistry and Human Nutrition, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76-100, and lDepartment of Food Science, Agricultural Research Organization, The Volcani Center, PO Box 6, Bet Dagan 20-250, Israel p-Vinyl-guaiacol (PVG) is one of the undesirable aroma components produced in orange juice during storage. The odor of PVG in orange juice can be detected by panelists at a concentration of 75 /ig/1 [Tatum.J.H. et al. (1975) /. Food Sci., 40, 707-709]. The aim of this investigation was to test the hypothesis that free ferulic acid is a precursor of the produced PVG. A method for the isolation and quantitative determination of free and of bound ferulic acid from orange juice was developed using t.l.c. and h.p.l.c. Results indicated that most of the ferulic acid in orange juice occurs in a bound form, undergoing some release during pasteurization and storage. Sensory evaluations, using the hedonic multiple-comparison procedure, revealed that orange juice, when incubated with added free ferulic acid, produced a significantly less acceptable aroma than juice incubated without the added ferulic acid. These changes in aroma quality could not, however, be detected by panelists during triangular or paired comparison tests. Changes in aroma quality and the increased level of free ferulic acid content during incubation were affected by time and temperature. It is concluded that although the content of free ferulic acid is low, it has the potential to cause a decline in aroma quality and this potential increases at elevated temperature and during long storage periods, and that this decline in aroma quality is concomitant with increased PVG content.

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ECRO VII: Abstracts

An automatic and ergonomic machine for odour theshold determinations


Steve Van Toller, David Sleight, Graham Wales and Jim Brockbank Department of Psychology, University of Warwick, Coventry, UK Because of the large ranges required, determination of a subject's odour threshold is usually not possible using an olfactometer. Use of a 1/3 or 2/5 forced-choice procedure means that the quantity of glassware required is very large. In addition, the glassware used to present the smell needs to be of a reasonably large size to allow for an adequate headspace above the odorous solution. A typical threshold determination will require upwards of 5 0 - 6 0 , 150 ml volumetric flasks. These will need careful washing to eliminate residual smells. Finally, there are a series of ergonomic problems relating to the need to ensure that the subject's select samples from the correct row, etc. An apparatus was designed to minimize such problems and allow threshold determinations to be made with the minimum of supervision. The apparatus designed consists of an odour Ferris wheel allowing for 10 concentrations of the odour to be tested. Each of the 10 rows contains five samples. The design uses a 2/5 force-choice, subjects are presented with a single ascending concentration row and they are required to select two of the five samples before being allowed to advance to the next concentration. Problems associated with clean glassware are overcome by using cheap disposable plastic cups with lids. The cups allow for a large headspace above the odorous solution. The apparatus is automatically controlled with a subject's selection for each row being entered into a computer. An updated collection of data, divided into sex and age can be displayed alongside the subject's own results.

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Memory for odours, colours and sound


Steve Van Toller and David Sleight Department of Psychology, University of Warwick, Coventry, UK Engen (1982) reported that it took subjects four times as long to name free recall of 10 odours as it did to name 10 colours. He concluded that odour language was idiosyncratic and impoverished. However, there are a number of factors that could explain his interesting finding without needing to suggest that odour is a 'special' sense. For example, colour names are usually taught to young children resulting in the fact that discrimination of colour will be easier and undertaken at a much finer level than that found for odour. It might also be argued that sound will be intermediate in that it is easier to recall the names of sound than it is to name odours but not so easy as it to name colours. Sixty two females and 62 males, were asked to name 10 colours, odours or sounds. The three items were counter-balanced into: Colours Sounds Odours Colours Sounds Odours Sounds Odours Colours Odours Colours Sound Odours Colours Sounds Sounds Odours Colours

Subjects were asked to name five items and then asked to name a second five items. Response times were recorded for sets of five recalled items. The overall average times of the freely recalled items were: Females Colours Sounds Odours 6.9 s 16.9 s 23.6 s Colours Sounds Odours Males 10.8 s 19.7 s 21.9 s

Detailed analysis of the results is presented together with a frequency list of the words generated for colours, sounds and odours.

Preliminary study of intensity coding in the olfactory system: electrophysiological approach in the frog
Patricia Viret and Andr6 Duchamp Labaratoire de Physiologie neurosensorielle, University Claude Bernard, F-69622 ViUeurbanne Cedex, France Odour intensity and quality are simultaneously coded by electrical activity of neuroreceptor cells and olfactory bulb neurones. The coding of odour quality at a constant intensity level has been previously investigated.

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ECRO VII: Abstracts Our aim is to study intensity coding performed by these two cell populations. Unitary extracellular activity was recorded in response to four odour stimuli: camphor, anisole, limonene and isoamyl acetate. The stimuli were delivered by a constant flow-rate olfactometer as 2 s pulses of 21 different concentration levels, the concentration range being 1 x 1 0 ~ ' - 7 x 10" 2 of the saturated vapour (at 22C). The present data allow us to draw dose response curves from a set of 48 neuroreceptor cells and 65 dose response curves from a set of 154 deutoneurones (these cells are more selective and thus, with four stimuli, the rate of noresponse is rather high). For instance, data processing led us to investigate, for each cell population and for each stimulus, the dynamics of the recruitment of the cells as a function of increasing concentration. This allows us to discuss inter-stimulus differences respectively at the periphery and in the olfactory bulb, and compare the sensitivity and the dynamics of the recruitment of the two cell populations, for each stimulus. Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on February 16, 2014

Olfactory organs in pelagic and benthic elasmobranchs


E.Zeiske, B.Theisen1 and H.Breucker2 Zoologisches Institul und Zoologisches Museum, Hamburg University, FRG, ^Institute of Cell Biology and Anatomy, Copenhagen University, Denmark and 1Anatomisches Instirut, Hamburg University, FRG The olfactory organs of elasmobranchs are either separated from the mouth or connected with it by nasolabial grooves concealed by nasal flaps. The former type is considered an adaptation in mostly pelagic living elasmobranchs in which the olfactory organs, during active swimming, are ventilated passively by a steady water flow. In the latter type, a pulsatile water flow through the organ is generated by respiratory activities and this ensures ventilation of the olfactory organs in mainly benthic living and often bottom resting species. Elasmobranchs show, however, very different evolutionary pathways in shape and position of their olfactory organs, even within smaller systematic entities. In order to demonstrate this variety, exemplified by the pelagic silky shark (Carcharhinus falciformis), the benthic small-spotted catshark (Scyliorhinus canicula) and the benthic or bentho-pelagic thornback ray (Raja clavata), a comparision is made between their olfactory organs. They are functionally divided into an inlet and an outlet chamber. The water enters the inlet chamber through the incurrent nostril and passes between olfactory lamellae lodging in microvillous receptor neurons before it is discharged via the outlet chamber and, finally, the excurrent nostril. The water flow through the organ is maintained by pressure differences between the nostrils achieved by forward motion in all three species, and also by coupling to respiratory movements in benthic species, as observed in resting thomback rays. Devices for putative pressure regulation, functioning as bypasses or shunts, are present in the three types of olfactory organs. Theisen.B., Zeiske.E. and Bruecker.H. (1986) Ada Zool. (Stockh.).

Histoenzymologic study of original cholinergic projections to the rat olfactory bulb: spatial distribution and ending sites
Li Mou Zheng, Nadine Ravel and Francois Jourdan Laboratoire de Physiologie neurosensorielle (UA CNRS 180), F-69622 VMeurbanne Cedex, France Serial sections have been realized in a region extending from the olfactory bulb to the magnocellular preoptic nucleus in order to study, by the AchE detection method, the projection sites of centrifugal cholinergic afferents to the olfactory bulb. In this study we demonstrated that the spatial distribution of cholinergic centrifugal fibers over the rat olfactory bulb is heterogeneous. Beyond the classically described staining in the internal plexiform and glomerular layers we observed dark spots of intense AchE labelling located at the border of the glomerular layer, close to the lateral olfactory tract and in the dorso-medial region, near the accessory olfactory bulb. These labelled areas, which disappeared after a section of the olfactory peduncle, coincide with anatomical units lying in close continuity with the glomerular layer. These glomerular-like structures are made of a dense neuropile surrounded by periglomerular cells. However they lack the nodules of densely packed olfactory terminals which characterize the typical glomeruli. Most of their synaptic terminals contain electron-dense vesicles, never observed in classical olfactory ones. The AchE-positive reaction, mainly intracellular, is found within small myelinated axons and large dendrites. These 'atypical glomeruli' could represent a peculiar site of convergence for original centropetal primary axons and centrifugal cholinergic fibers. The hypothesis of their implication, as a whole, in a new olfactory sub-system deserves further investigations.

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Goldfish olfactory bulb neurons respond to mechano-, thermo- and non-olfactory stimuli applied to the olfactory mucosa
H.P.Zippel, R.Voigt, T.Wachter and J.KQhling-Thees Physiologisches Institul der Universitat, Humboldtalle 23, D-3400 GOttingen, FRG Multunodality in the olfactory system and the distinction between taste and smell in aquatic vertebrates are controversial subjects. The latter because the stimuli reach both external chemical senses in the same medium. The present electrophysiological investigations were undertaken to test whether - in comparison with natural and synthetic olfactory stimuli - mechanical (drops of water, laminary water currents) and low temperature (local cooling of the olfactory mucosa, steps in 5C) stimuli on one hand and non-olfactory stimuli (taste, different salt concentrations, different pH values) on the other have a meaningful influence on single olfactory bulb neurons. The recordings were made on-line using a PDP-12 computer, and the effects (30 s stimulus application) calculated on the basis of the spontaneous activity (30 s preceding the stimulus) 2 5 % deviation from the spontaneous activity were (due to fluctuations in the spontaneous activity) taken as indifferent. On the basis of the computer analysis the stimuli can (on the basis of their effectivenss) be presented in a hierarchical order (% of responses > 25% in brackets): (l) natural odours: skin extract (71%) > Tubifex food extract (68% > rotten Tubifex extract (60%) > single amino acids (52%); (ii) mechanical stimuli: 30 s laminary current (69%) > long lasting laminary current (2 min); 52% of the neurons were found at a new 'spontaneous' level without any sign of adaptation > single drops of water (43%); (iii) local cooling: effects nearly linearly depending on the temperature step from the 'resting temperature' (23%): 3C step = 20C(22%)...23Cstep = 0C(64%); (iv) synthetic odours (46%): in contradiction to the above effects taste stimuli (sweet, bitter: 32%), salt solutions (NaCl 10" 2 , 10" 3 , 10" 4 molar: 30%) and pH values (pH 4, 5, 8: 42%) near or within the biological range elicited only weak effects. The pronounced effects recorded during application of organic acids (acetic, malomic acid: 69%) seems to depend mainly on their low pH (near 3), and are comparable with those obtained with pH 2 (66%). It is likely that the strong effects caused by a 1 M salt solution (61 %), and also by pH 2, merely represent reactions to inadequate stimuli outside the biological range. The above data demonstrate: (a) that the goldfish olfactory system is multimodal, but (b) is not equally sensitive to olfactory and gustatory stimuli.

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2. Taste Behavioral reactions to taste stimuli in hatchling chicks


A.Braun, J.R.Ganchrow and J.E.Steiner The Hebrew UniversityHadassah Faculty of Dental Medicine, Department of Oral Biology, Jerusalem, Israel Taste buds in chickens begin to develop ~4 days prior to hatching (Ganchrow and Ganchrow, 1986). Behavioral responses to taste stimuli dissolved in egg fluids have been observed in chick embryos 1 day prior to hatching (Vince, 1977), but not if water is the solvent. In contrast, adult chickens exhibit oral behaviors to chemical stimuli dissolved in water, but these give the impression of being mainly 'aversive responses' (Gentle, 1982). The present research investigated spontaneous behavioral displays of 28 chicks within 24 h after hatching. The free-moving chicks were presented with double-distilled water and aqueous taste solutions in a random sequence. Responses were videotaped and scored under double blind conditions. Specific movement features were counted for 1 min following the first detectable sampling. When compared to water, fewer drinking contacts were initiated when quinine (0.001 and 0.02 M) or citric acid (0.01 and 0.1 M) were given and increasing occurrence of features such as beak wiping, gapes and prolonged beak clapping bouts were observed. These latter behaviors together with head shaking appeared to aid in removing the 'offending' solution from the oral cavity. In contrast, responses to the 'sweet' stimuli (0.3 and 1.7 M fructose and 0.005 and 0.02 M sodium saccharin) were much less impressive and barely distinguishable from water reactions. Preliminary analyses indicate a tendency for fructose to elicit slightly more fluid contacts than water. These contacts were often followed by rapid beak opening and closing (clapping). For instance, only 22% of the animals clapped >20 times per contact when water was the stimulus, while 62% and 66% responded this way to 0.3 and 1.7 M fructose, respectively. Saccharin was intermediate (40% and 32% for 0.005 and 0.02 M, respectively). Hedonic estimates confirmed these results wherein the higher concentrations of quinine and acid were rated as clearly unpleasant ( - 1 . 5 on a 10 cm visual analogue scale) while the 'sweet' and water responses hovered around neutrality (the higher concentrations being slightly more positive). It is concluded that hatchling chicks display an adult-like array of reactions to taste stimuli. The

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ECRO VD: Abstracts minimal positive reaction to the sweet stimuli is perhaps related to preference findings from some laboratories suggesting chickens are indifferent to these substances. This research was supported by grant number 3226/84 from the United States-Israel Binational Science Foundation (BSF), Jerusalem, Israel.

The four basic tastes test


L.J.van Gemert TNO-CIVO Food Analysis Institute, PO Box 360, 3700 AJ Zeist, The Netherlands From a review of the relevant literature it was concluded that in selecting or screening assessors of sensory panels for the four basic tastes test different compounds and especially different concentrations (with ratios up to 40) are applied. Furthermore, results are interpreted in different ways. In order to check on reproducibility and valid concentrations we performed an experiment in which 35 subjects (12 women, 23 men, age 2 3 - 5 2 years) tasted five items each with an interval of 1-2 months, 14 stimuli [three different concentrations in water of: sucrose (4, 8, 16 g/1), citric acid monohydrate (0.2, 0.4, 0.8 g/1), sodium chloride (0.5, 1, 2 g/1) and caffeine (0.25, 0.5, 1 g/1) and two blanks]. The scores were formed by the number of correctly identified stimuli. The test appeared to be moderately reproducible; the scores ranged from 4 to 14 with a maximum difference per subject of 6. All concentrations showed percentages correct ranging from 40 to 90, except sucrose at 16 g/1 for which the percentage correct appeared to be nearly 100. Correlation with results of triangle tests (with a large diversity of food products and differences involved) shows that subjects with a low score on the four basic tastes test also have a low percentage of correct triangle tests. Subjects with a high score show high as well as low percentages of correct triangle tests. Based on the results obtained, a simplified testing method is proposed. First of all, five stimuli (sucrose 10 g/1, citric acid monohydrate 0.8 g/1, sodium chloride 2 g/1, caffeine 1 g/1 and a blank) are presented labeled to make the subjects acquainted with the tastes involved. Then 10 coded stimuli are served in which each of the five stimuli occurs twice in random order. If all responses are correct, the subjects will be allowed to take part in subsequent tests. If not, the test will be executed one more time (after discussing the incorrect responses).

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Beidler's taste mixture equation tested psychophysically with glucose-fructose mixtures


Cees de Graaf and Jan E.R.Frijters Department of Human Nutrition, Netherlands Agricultural University, Wagtningen, The Netherlands Beidler's taste mixture equation describes the relationship between the concentration and composition of a binary mixture and the magnitude of the neural response. It aims to describe the peripheral taste interaction between two taste substances in a mixture under the condition that the molecules of both substances compete for the same receptor sites. The purpose of the present study is to show that Beidler's mixture equation can be tested appropriately with indirect psychophysical methods, without the necessity of making assumptions about the magnitude of the maximum responses to the single compounds which constitute the mixture. Also, the values of the association constants need not to be known. An essential condition for such a psychophysical test is that both substances have similar tastes. In a series of experiments, fructose and three equiratio mixtures of glucose and fructose were matched in perceived sweetness intensities to five different glucose concentrations using the method of constant stimuli. The results showed that Beidler's mixture equation adequately describes the taste interaction between glucose and fructose at low sweetness levels. Concentrations predicted by the mixture equation deviated on average 2% from the concentrations experimentally determined. At high sweetness levels the taste system is more efficient than expected on the basis of the mixture equation, because the mean mixture concentrations experimentally determined were ~ 9 % lower than those predicted. These findings suggest that glucose and fructose share common sites, but either one or both have additional secondary binding mechanisms.

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ECRO VII: Abstracts

Selective fluorescent marking of fish taste buds damaged by detergents


A.Holl Justus Liebig-UniversilOt Zoologisches Institul, D-63 Giessen, FRG Histologies] and cytological examination of the barbels of the tench (Tinea tinea) and of the bullhead (lctalurus nebulosus) previously exposed to sublethal concentrations (0.055.0 p.p.m. for up to 25 days) of Persil reveals delicate deleterious effects to central taste bud structures. Such inconspicuous 'early' signs of histological or cytological damage are difficult to recognize when employing routine staining procedures. By intravital application of the fluorescent dye Procion Brillant Yellow to the surface of the affected barbels damaged cells become quite distinctly visible. Selective fluorescence is due to the uptake of the Procion dye only by disintegrated cells. The staining procedure and preparation for fluorescence microscopy is described. The method may also be well suited to the recognition of sublethal taste bud damage caused by substances other than detergents. Supported by DFG.

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Multisapophoric molecules
S.Shami] and G.G.Birch Department of Food Technology, Food Studies, University of Reading, Whiteknights, Reading, Berks RG6 2AP, UK Multisapophoric molecules may possibly be defined as molecules known to generate more than one type of taste. Other types of molecule may possess two or more potential sapophores yet generate only one type of taste. As well as possessing the appropriate sapophore, a molecule must accede to an appropriate region of the taste epithelium for a particular taste to be generated. This involves the solution thermodynamics of the sapid solute and its compatibility with water structure.

The sensory determination of equi-sweet concentrations of certain sweeteners


Anne Tunaley, D.M.H.Thomson and Jean A.McEwan Department of Food Science. University of Reading, PO Box 226, Whiteknights, Reading RG6 2AP, UK Qualitative and quantitative comparisons of the sensory characteristics of swectners require that they should have approximately the same sweetness intensity. To this end, the concentrations were determined at which aqueous solutions of fructose, glucose, sorbitol, lactitol, aspartame, saccharine and acesulfame K, were perceived as equi-sweet to a 5% sucrose standard. Pilot studies were used to identify a range of three concentrations of each sweetener, such that the corresponding range of sweetness intensities included that equivalent to the sucrose standard. The sweetness intensity of each concentraton of each sweetener was then rated in comparison to 5% sucrose. Sweetness ratings were made on a 150 mm continuous line scale, anchored at the left extremity, middle, and right extremity, with 'much less sweet', 'standard' and 'much more sweet' respectively. This type of scale had not previously been empoyed in this context. The experimental design ensured that each sweetener appeared with every other sweetner, and sample presentation was randomised within and between assessors. At each experimental session, the 31 assessors tasted the standard and six samples; the experiment was replicated three times. The data was analysed using GLIM and SAS computer packages. It was evident from the differences within and across individuals, that it is more appropriate to consider an equi-sweet range for each sweetner, rather than a single concentration. However, future practical considerations necessitate the use of consensus equi-sweet concentrations. Variability of results was also sweetener dependent; the more complex the total perception associated with a sweetener, the more variable the results.

Afferent connections of the caudolateral orbitofrontal cortex taste area of the primate
Leslie L.Wiggins, Edmund T.Rolls and Gordon C.Baylis University of Oxford, Department of Experimental Psychology, Oxford 0X1 3UD, UK A cortical taste region has recently been identified in the caudo-lateral orbitofrontal cortex of the primate (1). In this area, single neurons are sharply tuned to gustatory stimuli, and are influenced by the motivational state of the monkey. In order to determine the afferents to this region of taste cortex, and to determine whether

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ECRO VII: Abstracts his a primary, secondary or tertiary region of cortical taste processing, injections of WGA-HRP for retrograde ueuronal tracing were made into this region in three monkeys in which the exact location of this cortical taste region had been identified by recordings of the activity of single neurons. Labelled cell bodies were found in the frontal opercular taste cortex and in the insular taste cortex, both of which are primary taste cortices in that they receive from the thalamic taste nucleus, VPMpc. Further, the caudolateral orbitofrontal cortex taste area did not receive inputs from VPMpc, but instead received projections from the mediodorsal nucleus of the thalamus, the thalamic nucleus which projects to the prefrontal cortex. These results show that the caudolateral orbitofrontal taste cortex is a secondary taste cortical area, and that it receives gustatory inputs from the frontal opercular and insular taste cortices. Afferents were also shown to reach the caudolateral orbitofrontal taste cortex from the more ventral part of the rostral insular cortex, the amygdala, the substantia innominate, the rhinal sulcus and from the surrounding orbitofrontal cortex. Through some of these pathways visceral information important in modulation of responsiveness by motivational state may reach the caudolateral orbitofrontal taste cortex. 1. Rolls.E.T., Yaxley.S., Sienkiewicz,Z.J. and Scott.T.R. (1985) Chem. Senses, 10, 443.

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Time intensity curves of taste (2)


M.Yoshida Chuo University, Bunkyo-ku, Tokyo Yoshida (1986) and Yoshida and Mochizuki (1985a,b) reported a microcomputer system to record and analyze time intensity curves for some natural and synthetic sweetncrs. Here, with the same technique as above, time intensity curves for alanine, glycine, stevioside and palatinose were recorded. Concentrations were set to be equivalent with sucrose 2.5, 5.0 and 10.0%. Subjects were six male students. (In the preliminary studies > 14 students participated, however, a few were withdrawn due to inconvenience, while the rest were withdrawn due to lower reliability, larger variance at every second.) The results were very similar to those reported in Yoshida (1986). Training effect or convergence of variance was also traced (average for the first trial, for the second trial, etc.). A few minor modifications were added to the programs. (Since some of the comments were translated into Japanese, Western researchers may regard them as a kind of regression.)

3. Chemoreception in Invertebrates Processing of the odorant information in the honeybee: a functional and immunocytochemical study
Ge'rard Arnold, Claudine Masson and Jean-Pierre Denizot1 CNRS-INFA, F-91440 Bures sur Yvette, and 'CNRS, F-91190 Gif sur Yvtlte, France In the glomenili of the antennal lobe, the olfactory messages undergo an important integration and encoding before being transmitted to the higher centers. The neural mechanisms underlying this integration are analysed on the one hand with the method of 2-deoxyglucose (2-DG) autoradiography, and on the other hand, with an immunocytochemical study. The 2-DG method is used to study the role of spatial factors in olfactory processing. Under controlled odor conditions (e.g. isoamyl acetate), there is a characteristic pattern of selective glomeralar 2-DG uptake; a correlation between the quality of the odor and the pattern of glomerular activity has to be confirmed with other compounds. Immunocytochemical study shows that GABA and its biosynthetic enzyme GAD are abundant in the antennal lobe. GABA immunoreactivity is mainly found in neuronal cell bodies and in their processes in the neuropile, whereas GAD immunoreactivity is found in the cortical layer of the glomeruli where the synaptic connections are localized. These results thus support the hypothesis that inhibitory interneurones have a crucial function in the processing of the odorant information at this first sensory level of the central nervous system. Furthermore, at the higher level of integration, as for example in the calyx of the mushroom bodies, our results show that some endings of efferent deutoneurones are also GABAergic.

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Antenna! sexual dimorphism in the distribution of gustatory and olfactory sense organs in Blaberus craniifer (Insecta, Dictyoptera)
I.Chambille Laborawire de Neurobiologie Compare des Invertibrts, CNRS/INRA. F-91400, Bures sur Yvette, France The morphometiy and the distribution of four types of chemoreceptors are studied at different stages of development. The smallest sensilla consist of three classes (8, 12 and 20 jtm in mean length). According to their structural characteristics and first electrophysiological results (1) an olfactory function can be assigned to them. The contact chemoreceptors belong to the fourth class, these are long mobile bristles (length: 22-85 ^m) well-ordered around the antenna] segment, with a very grooved wall and a terminal pore (tp-sensilla, 2). They are 20% of the whole chemosensory population at the 1st stage (jc = 450) and only 10% in adult (x = 5200). A strict agreement with Leucophaea (3) and Periplancta (4), antennal sensilla can be deduced while examining by light and scanning electron microscopy. For the same number of flagellar annuli (n = 26; 1st instar), male nymphs bear on the flagellum more gustatory (total difference d = 4%) and olfactory (d = 11%) sensilla than females. The distributions are statistically significantly different for all the flagellum for olfactory hairs (particularly those of the smallest) and only for the 21 proximal annuli for gustatory hairs. During nymphal instars, the number of flagellar annuli rises with the same rate in each sex and gustatory dimorphism always increases (the olfactory one is being worked out). In adults, males show on average 750 gustatory receptors more than females (n = 103). This difference (d = 16) results from a significantly more important number of sensilla on each segment wherever the antennal portion is considered (with poor equipment as proximal part: 11 sensilla/segment or with a rich one as median part: 65 sensilla/segment). In various species of cockroaches, the sexual behaviour consists of a series of sequences, in most cases, released by chemical and tactile stimuli received by antenna] sense organs. In males of Blaberus, the functional implication of the olfactory and gustatory/tactile dimorphism must be sought in the mating (perception at distance of mature females and release of courtship activities) as well as in their more specific behaviours [gregarism (5) and aggressiveness]. 1. 2. 3. 4. 5. Lambin.M. (1973) These de 3me cycle, University de Dijon. Altner.H. (1977) Olfaction and Taste VI. Paris, pp. 295-303. Schaller,L. (1982) Cell Tissue Res., 225, 129-142. Schaller.D. (1978) Cell Tissue Res., 191, 121-139. Brossut.R (1970) C.R. Acad. Sci., Paris, 270, 714-176.

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Taste and olfactory responses of ovipositor chemosensilla in Rhagoletis pomonella


R.Crnjar, A.M.Angioy, J.G.Stoffolano, Jr 1 , I.Tomassini Barbarossa and P.Pietra2 Institute of General Physiology, University of Cagliari, Italy, ^Department of Entomology, University of Massachusetts, Amhent, MA, USA, and 1Institule of Human Physiology, University of Modena, Italy In the apple maggot fly, RhadoUtis pomonella, four pairs of chemosensilla are present in the sub-apical region of the ovipositor and all but one pair are located within two lateral grooves. This study aims to provide information on the sensitivity and discriminative properties of these sensilla in response to substances present in, or related to, oviposition substrates (apples). With different electrophysiological techniques, spike discharges were recorded following contact stimulation (taste experiments) and slow compound potentials (EOvGs) were recorded following stimulation with volatiles (olfactory experiments) from same chemosensilla. In taste experiments, analysis of spike discharges showed that: (i) solutions of NaCl, glucose, fructose, malic and quinic acids always elicit polyneuronal responses ( 3 - 4 spike types); (ii) malic and quinic acids often evoke volleying activity in the tonic portion of the discharges; (iii) response patterns to glucose, malic and quinic acids differ from that of NaCl; the response pattern to glucose is different from those of malic and quinic acids; no differences are found between responses to NaCl and fructose or between those of malic and quinic acids. Other compounds tested are not stimulatory. In olfactory experiments, the following stimuli elicit, in the order of EOvGs of decreasing mean amplitudes: phenol, ethanol, isoamyl acetate, methyl eugenol, apple extract, pyrocatechol, oxidized apple odor. All other substances evoke responses of little or no importance. The ovipositor chemosensilla appear to be sensitive to volatile and contact stimuli present in apples (oviposition substrate) and may provide the egg-laying fly with discriminative information among the latter.

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Preliminary results on the neuronal bases of odour memorization by gustatory reinforcement in the honeybee Apis mellifera
Caroline Fonta and Claudine Masson INRA-CNRS, F-9144O Bures sur Yvette, France The location of food sources by honeybees relies upon the memorization of sensory stimuli. The association between the floral aroma (conditional stimulus, CS) and the food reward (inconditional stimulis, IS, and reinforcement) is particularly effective in eliciting conditioned feeding behaviours (1). In order to analyse the neuronal bases of this bisensory (olfactory + gustatory) integration, we first determined the nervous pathways involved in CS and IS processing, in the central nervous system. We studied the structures which receive inputs both from antennae (CS) and from mouthparts (IS) by using cellular staining methods with Co2"1" and Ni 2+ ions. The suboesophagcal ganglion is the first level of projection for mouthparts fibres. Tongue fibres penetrate into the anterior lateral part of the antennal lobe; the projection area corresponds to the anterior and dorsal group of deutocerebral glomeruli (2) which receive inputs from the antennae. The gustatory fibres do not have many ramifications and exhibit varicosities. These preliminary anatomical data suggest a possible interaction between antennal and buccal inputs below the upper brain centres (such as the protocerebrum known to be the multisensory integration level), in the deutocerebrum. Functional studies are in progress to analyse the temporal parameters of gustatory inputs on the central processing of olfactory signals. Extra- and intracellular recordings coupled with dye injection are carried out in the deutocerebral cell population. Analyses at this first convergence level of gustatory and olfactory inputs might partly explain the initial steps of the reflex proboscis extension and of the olfactory mnesic trace. 1. Menzel.R. (1967) Untersuchungen zum Erlernen von Spektralfarben durch die Honigbiene (Apis mellifica). Z Vergl. Physiol, 56, 2 2 - 6 2 . 2. Arnold,G., Masson,C. and Budharugsa.S. (1985) Comparative study of the antennal lobes and their afferent pathway in the workerbee and the drone (Apis mellifera). Cell. Tissue Res., 242, 593-605.

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Morphofunctional and behavioural evidence for a critical period in the development of the olfactory system of honey bee
J.Gascuel, M.H.Pham-Delegue, C.Masson and G.Arnold INRA-CNRS, F-91440 Bures sur Yvette, France In the honey bee, anatomical and electrophysiological data have revealed a maturation in the setting up of glomeruli (where synaptic connection between afferent olfactory neurons and deutoneurons occurs) and in antennal electrophysiological responses (EAG), from 3 days before emergence to 8 days of adult life. From this basic work, in order to define limits of bee olfactory plasticity, we carried out multidisciplinary approaches to determine epigenetic effects on the structure and function of the olfactory system. At the structural level, quantitative analysis by a semi-automatic measurement of histological sections (obtained after cobalt ion migration) showed that rearing in low odour conditions during the nymphal stage and the first days of adult life induced a decrease in the cortical layer volume of glomeruli and in the whole volume of each ordinary glomerulus. Moreover, such conditions of rearing led to a lower level of electrophysiological (EAG) responses. At the behavioural level, a biological test was set up, using a four armed airflow olfactometer, in order to record, by means of video, the olfactory choices of individual worker bees. Bees previously submitted to various periods of olfactory imprinting showed significant differences in later responses towards the imprinting scent, compared to naive bees. Current experiments should determine how long such behavioural changing will last, compared with classical olfactory conditioning. These convergent data will allow determination of the hypothesis of a critical period, occurring from 3 days before emergence to the first days of adult life, where olfactory experience is decisive in the later functioning of the honey bee olfactory system and final stabilization of synaptic connections.

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A method for correlation of electrophysiology and distribution patterns of pheromone-sensitive sensilla in a moth
Eric Hallberg and Bill S.Hansson University of Lund, Sweden To achieve a positive identification of pheromone-sensitive sensilla trichodea in Agrotis segetum the following procedure was performed. (i) Recording of the sensilla response using the tip recording method. The pheromone substances used included Z5-10:OAc, Z7-12:OAc and Z9-14:OAc. The responses were measured from sensilla on specific segments (no. 15, 25 and 35), on the lateral or medial halves of the antennae. (ii) To mark the segment, one of the terminal sensilla chaetica was broken using glass knives. (iii) The antennae were fixed in 70% ethanol, air-dried, mounted on holders and sputtered with gold/palladium. (iv) In the scanning electron microscope it is possible to find the electrophysiologically defined sensilla using the following steps: counting the segments from the base to get a rough identification of the segment; looking for the broken sensillum chaetica to get a positive identification of the actual segment; finally the sensillum from which the recordings were made can be found because the tip of the pheromone-sensitive sensillum has been broken before the recording.

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Function and development of sensory cells in 'attractant sensilla': physiological differences between larvae and adults of Periplaneta americana L.
Norbert Hartmann Institut fir Zoologie, Universitdt Regensburg, FRG The most common olfactory type of sensillum on the antennae of adult male P. americana is a 4-celled, single-walled sensillum (long type-swB, 25 ym) with receptor cells for female sexual attractant (1,2). Schaller's study (1) showed that the number of these sensilla doubles during the imaginal moult; half of these develop from extensions out of the 4-celled short sensilla (short-type-swB, 15 ^m) found in male nymphs. As was shown by electroantennograms, the male nymphs are already sensitive to the components of the female attractant (3). The question therefore arises as to whether the sensory cells of these morphologically different sensilla have different physiological characetristics. For this reason the corresponding sensilla of male nymphs and adults were examined electrophysiologically. Single cells could be distinguished. The effect of the components of the female attractant and ~60 other volatile substances, mainly from foodstuffs, were tested on them. In adult males two cells respond to the substances in the female attractant, one not only to periplanone B but also to periplanone A as well as other unidentified components in extracts of the attractant. The other cell is sensitive to periplanone A and two other behaviorally active substances. These are B- and A-cells (2, and Sass and Hartmann, in preparation). Pheromone receptor cells of male nymphs are 100 times less sensitive to periplanone A and B than those of the adults. The swB senilla of both stages also have a third and a fourth cell. These respond to terpenes, alcohols, amines, and aromatic compounds. Characteristic curves from 1,8-cineol and (+)-fenchone fail to show a significant difference in sensitivity between male nymphs and male adults. 1. Schaller.D. (1978) Cell Tissue Res., 191, 121-139. 2. Sass.H. (1983) /. Comp. Physiol., 152, 309-317. 3. Nishino,C. and Kimura.R. (1982) Comp. Biochem. Physiol., 72A, 237-242.

Sensory equipment of the 17-year cicada: structural survey and physiological evidence for olfaction
Ulla Klein, Cornelia Bock, Wolf A.Kafka and Thomas E.Moore1 Max-Planck-Institul fUr Verhaltensphysiologie, D-8131 Seewiesen FRG, and 'Museum of Zoology, University of Michigan, Ann Arbor, Ml, USA The periodical cicadas of North America are famous for their outbreaks every 17th year at one geographical place. Maier [Ann. Entomol. Soc. Am., 75, 14 (1982)] reported that females oviposited on 49 species of

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ECRO VII: Abstracts woody plants among which only 33 species were accepted for feeding. Very probably, chemical stimuli are involved in feeding and oviposition, and might thus contribute to the mechanisms by which the limited resources, e.g. of egg laying places, are shared. For a better understanding of the breeding ecology and the spatial dynamics of the dense cicada population a deeper knowledge of the chemosensory inputs guiding individual behaviour of host plant selection is necessary. With scanning electron microscopy a search for possible candidates for chemoreceptive sensilla was started at the beak, the ovipositor and the antennae of Magicicada cassini (Fisher) (Homoptera, Cicadidae). On the tip of the labium, a ventral and a lateral group of sensilla were conspicuous implying special functional units. On the ovipositor, rows of small dome-shaped sensilla embedded in the cuticle surface were found proximal and distal on the first and second valvulae. The antennae, similar in both sexes, consisted of scape, pedicel and a 5-segmented flagellum. Most of the flagellum lacked almost any seta. The ventral side of the first segment, however, was covered by a dense field of sensilla coeloconica comprising - 6 0 large, 10 medium sized and 35 small sensilla. Transmission electron microscopy of the large s. coeloconica revealed multiporous single-walled pegs with pore tubules, housing three sense cells with highly branched dendrites. The small s. coeloconica were no-pore sensilla with inflexible sockets housing the 'triad' of dendrites, typical for hygro- and thermoreceptor-cell combinations. Electrophysiological recordings from single sensilla of the first flagellar segment showed specific reactions to cyclic terpenoids, constituents of plants that are usually not accepted as hosts. In accordance with the morphological findings about the large s. coeloconica, 2 - 3 sense cells could be distinguished with different but highly overlapping reaction spectra. So far, no responses to CO2, temperature or humidity were found.

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SensiUum lymph proteins from insect olfactory hairs


Ulla Klein, Maria Kehl1, Jflrgen Hemberger and Anka GOnzel Gruppe Kaissling, Max-Planck-Institut fUr Verhailensphysiologie, D-8131 Seewiesen, and iGenzentrum, MaxPlanck-Institut far Biochemie, D-8033 Maninsricd, FRG Two soluble proteins were found in antennae of male Antheraea polyphemus (Saturniidae), the 'pheromonebinding protein' (PBP) and the pheromone-degrading 'sensillar esterase'. They occur in the sensillum lymph of olfactory hairs and very likely play an important role in transduction. The PBP functions either as carrier [Vogt and Riddiford (1985) Proc. Natl. Acad. Sri. USA, 82, 8827] or, more likely, as inactivator of the stimulus molecule [Kaissling (1986) Anna. Rev. Neurosa., 9, 121]. Micro-electrophoretic analysis (ultrathinlayer isoelectric focusing and microgradient electrophoresis in polyacrylamide gels) of different preparations of sensillum lymph confirmed the localization and showed further molecular properties. The PBP and the sensillar esterase were detected only in preparations of male antennae. The PBP was the only protein which bound the tritiated pheromone in fluorograms. Glycoprotein staining revealed <0.5% sugar in the PBP, i.e. it is a non-glycosylated protein. In microgradient-gel electrophoresis, the PBP appeared as a double band, even in preparations of individual animals. The relative densities of the slow and fast band were 2 : 1 . Isoelectric focusing revealed a pi = 4.7 for the PBP and a pi = 3.0 for the sensilla esterase. The sensillar esterase had a molecular mass of 55 K. Further (integumental) esterases, found outside the olfactory hairs in male and female antennae, showed one main band at pi = 5.0, corresponding to the esterase with a molecular mass of 90 K, and two faint bands at pi = 5.85 and pi = 6.0, corresponding to the esterases around 65 K. The amino acid composition of the purified PBP was determined as 10 Ala, 2 Arg, 17 Asx, 6 Cys (1/2), 15 Gly, 5 Glx, 6 His, 4 lie, 9 Leu, 7 Lys, &3 Met, 5 Phe, 4 Pro, a: 17 Ser, 2:8 Thr, a 5 Tyr, 5 Val, Trp not determined (total number > 128). The molecular mass according to this analysis is 13.4 K. The first 16 NHj-terminal amino acids were determined by microsequencing: N-Tyr-Pro-Glu-Ile-Met-Lys-AsnLeu-Ser-Asn-Asn-Phe-Gly-Lys-Ala-Met-, in accordance with the result of Vogt and Prestwich (abstract, presented at ISOT IX, 1986) except for N-Ser at the start.

ATP and cAMP influence chemoreception in the blowfly


A.Liscia, R.Cmjar, A.M.Angioy, I.Tomassini Barbarossa and P.Pietra1 Institute of General Physiology, University ofCagliari, and lInstitute of Human Physiology, University of Modena, Italy Addition of ATP to stimulating solutions is known to lower the responses of labellar chemoreceptors of Phorma regina M. to NaCl and fructose and to enhance the responses to glucose and sucrose. On the other hand,

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ECRO VII: Abstracts cAMP addition decreases the responses to NaCl, fructose, glucose or sucrose solutions. The present study aims to provide further data on the interaction between ATP and cAMP (added to the stimulating solutions) and taste chemoreception in the blowfly. Results were obtained by means of the tip recording technique and show that: (i) the inhibitory effect of ATP on NaCl or fructose responses is suppressed by adding to the stimulating solutions an inhibitor of adenylate cyclase and thus of cAMP synthesis (GDP^S; that proved to be not stimulating 'per se"); (ii) addition of GDP/3S to glucose+ATP or sucrose + ATP solutions further enhances the increase in response due to ATP; (iii) GDP/3S added to NaCl (nucleotide free) solution enhances the salt response; all results, and particularly the last one, point to the existence of an adenylate cyclase activity between the sensillum tip and the receptor membrane. These findings suggest that: (a) cAMP (possibly synthesized from exogenous ATP) seems responsible for the inhibitory effect of ATP on NaCl or fructose responses; (b) in spite of the observed increase due to ATP on the responses to glucose and sucrose [possibly explained by an involvement of ATP in sugar reception mechanism(s)], cAMP synthesis from exogenous ATP applies also in this case. Finally, an hyperpolarizing effect of cAMP on chemoreceptor membranes, as previously reported for post-synaptic areas, becomes an attractive hypothesis to explain the reported inhibitory effect of this nucleotide.

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Adaptation and disadaptation of blowfly taste receptor responses and models of taste hair functioning
F.W.Maes, M.J.van den Berg and A.J.Schilstra Department of Animal Physiology, State University Groningen, PO Box 14, 97500 AA Haren, The Netherlands A chain of events links the application of a gustatory stimulus to the tip of an insect taste hair with the spike trains elicited from the receptor cells in the hair. These include: (i) diffusion of stimulus substance into the 'tip chamber' of the hair, where the ciliary tips of the taste receptors end, and removal of stimulus substance from this space; (ii) the primary process to taste receptor stimulation, presumably located in the membrane of the ciliary up; (iii) transmission of cellular excitation to the taste cell body; (iv) generation of action potentials. The functioning of each of these steps is far from clear, despite extensive experimental work done over the past three decades. In this contribution, the four steps are briefly reviewed, a number of possibilities, especially for the first and fourth step, are selected to be included in a model of taste hair functioning. Where specific information is lacking, general notions on receptor functioning are used. For instance, the fast adaptation of the spike rate to a tonic level, which occurs in - 1 5 0 ms, is ascribed to the fourth step. The models are computerfitted to a set of adaptation data of salt receptor responses from labellar taste hairs of the blowfly Calliphora vicina to 1 M KC1 (obtained by Maes and Harms, this volume), yielding a set of parameter values for each model. These are then used to predict the time course of disadaptation (recovery of responsiveness after cessation of stimulation). Experimental disadaptation data were obtained for adaptation times of 80 ms5 s and disadaptation times of 20 m s - 2 0 s, from 50 labellar salt receptors. Comparison with the predictions reveals that the first step, diffusion of stimulus into and removal from the tip chamber, is the critical one. Removal or inactivation of stimulus substance should occur at a rather high rate, indicating the presence of a fast, active removal process. The structural basis for such a process is discussed.

A study of plant chemical blends involved in honey bee foraging behaviour


M.H.Pham-Delegue, C.Masson, P.Etievant1, R.Marilleau and M.Le Metayer INRA-CNRS, F-91140 Bures sur Yvette, and lINRA, F-2100 Dijon, France Honey bee-plant relationships are based on a conditioning process where olfactory cues (plant aroma) and gustatory cues (mainly nectars) are closely linked and may lead to a selective foraging behaviour. In sunflower hybrid seed production, such selectivity may induce a lack of seed yield. In order to define the role and the nature of plant chemicals implied in pollination processes, we set up a study by combined behavioural and chemical assays. Studies in the field included the following, (i) Studies were carried out on two couples of genotypes producing different levels of hybrid seed yield (low yields for hybrid named Marianne, high yields for hybrid named Mirasol). Behavioural observations in controlled conditions of pollination under tunnels allowed us to point out a selective distribution of foragers among Marianne parental lines, while Mirasol parental lines

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ECRO VII: Abstracts were randomly visited, (ii) Volatiles emitted by living sunflower heads were trapped by the 'head-space' method; statistical analysis of chromatographic profiles obtained by gas chromatography (g.c), showed significant differences among the coupled genotypes, restricted to a few compounds (10% of the indexed compounds), (iii) Nectar samples collected with capillary glass tubes were analysed by g.c. Significant differences appeared among glucidic profiles of the different genotypes (particularly among sucrose amounts), related to genotype attractiveness. The following laboratory studies were carried out. (i) The foraging situation is simulated in a flight room, using an artifical flower device for a biotest based on olfactory conditioning. Behavioural responses combined with chemical fractioning of sunflower solvent extract, showed that honeybees may recognize sunflower aroma from a limited range of component (10% of the whole blend), partly identified by g.c.-m.s. (ii) Gustatory preferences among sugar consituents of nectar were evaluated after a gustatory conditioning. Foragers elicited preferences for the highest concentrations (>50%) of each sugar (sucrose, glucose, fructose) and for sucrose among the different sugars at their optimal concentration. Moreover, the recruiting process, which is a basic phenomenon of foraging behaviour, was the highest with sucrose solution. Thus, it appeared that even though volatile blends were much more complex compared to glucidic blends, for both kinds of cues, only a 'limited chemical pattern' was responsible for foragers' choices.

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A morpho-functional investigation on the antennal olfactory receptors in the medfly, Ceratisis capitata Wied
G.Scalera, A.Bigiani, R.Crnjar1 and P.Pietra Istituto di Fisiologia Umana dcll'Universita di Modena, and llstituto di Fisologia Generate dell'Unviersita di Caglairi, Italy Information was collected regarding the typology and topology of sensilla with a probable olfactory function sparsed on the surface of the antennal flagella in Ceratitis capitata. These data were compared with EAG recordings performed on the same sensilla stimulated with various volatile compounds (mostly alcohols and aldehydes C2CIO). Both the morphological and EAG data were grouped according to eight equivalent sectors into which the flagellar surface had been arbitrarily divided. The morphological data, obtained from S.E.M. pictures, showed that three types of sensilla probably housing olfactory receptors were present on the antennal flagella: long basiconica (B), trichodea (T) and short grooved basiconica (G). One single pit, containing two or more G sensilla, was present on the proximal portion of the lateral face; B sensilla were the most numerous and were most dense in the proximal and central sectors of the flagellum and around the pit; G sensilla were homogeneously scattered all over the flagellar surface; T sensilla were almost absent in the proximal portion but their density increased towards the flagellar tip. For most tested compounds, EAG amplitude varied as a function of the recording location. In fact, EAG amplitude was found to increase from proximal to central sectors - where it reaches its maximum and to decay steeply towards the distal sectors. Butanol, n-amyl alcohol, ethanol, 1-hexanol, hexanal, were stimulating in decreasing order, whereas 1-nonanol, 1-decanol and nonyl aldehyde had little effect. For each flagellar sector EAG amplitude values were also correlated to density values of B, G and T sensilla.

Electrophoretic pattern of soluble antennal proteins from some Lepidoptera


A.Topazzini, M.Mazza and P.Pelosi Istituto di Industrie Agrarie, Via S.Michele 4, 56100 Pisa, Italy As part of our research aimed at the identification of specific proteins involved in the perception of pheromones in insects, we report a preliminary mapping of soluble proteins in the antennae and other parts of the body of some Lepidoptera. The species studied were Bombyx mori, Lymantria dispar, Amata phegea, Pieris rapae and Melanargia galatea: for each of them we examined male and female antennae, head, thorax, abdomen, legs and wings. The crude extracts were analysed by SDS electrophoresis, isoelectric focusing in ampholine gradient and two-dimensional electrophoresis. The SDS gels usually show a most intense band, corresponding to a molecular mass of 18-20 led, which, in some cases, accounts for - 8 0 % of the total. The separations in ampholine gradients show a different quantitative distribution, suggesting that the major band in the SDS gel is made up of, or derives from, several different proteins.

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ECRO VII: Abstracts Therefore, the extracts were also separated on two-dimensional gels, using isoelectric focusing in the first dimension and SDS in the second. A comparison of the maps relative to the different parts of the body indicates the presence of some proteins specific for the male antennae. These proteins may have some role in the perception of the specific pheromone and are the most suitable for binding experiments.

Wind tunnel studies on an attraction-inhibitor in Coleophora laricella


P.Witzgall Max-Planck-lnstitut JUr Vernaltensphysiologie, Abt. Kaissling, D-8131 Seewiesen, FRG In previous field trapping tests, (Z)-5-decen-1-ol (Z5-10:OH) has proved highly attractive to C. laricella male moths, whereas the corresponding acetate (Z)-5-decen-l-yl acetate (Z5-10:Ac) acted as a strong attractioninhibitor (1). In the field, the attractant alcohol, which has recently been identified from female gland extracts (2), elicited upwind orientation flights, landing and walking near the pheromone source (3). These behavioural responses were strongly modified by the acetate inhibitor (4). The results of these field observations have now been confirmed by wind tunnel studies. Upwind flight tracks and the behaviour near the pheromone source were modified by an admixture of 0.01 % Z5-10:Ac. Blend sources containing up to 1% inhibitor still elicited some orientation flights, blends containing 10% acetate were not attractive at all. Tests with separate sources of attractant and inhibitor showed that orientation flights towards Z5-10:OH could not be fully suppressed when the same amount of Z-10: Ac was placed 5 cm apart. Here again, flight tracks differed characteristically from those obtained with pure attractant alone. 1. Priesner et al. (1982) Z Naiurforsch., 37c, 953-966. 2. Witzgall et al., in preparation. 3. Witzgall and Preisner (1984) Z Ang. Entomol., 98, 15-33. 4. Priesner and Witzgall (1984) Z Ang. Entomol., 98, 118-135.

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