1 Epitoky: Methods, Mysteries, and Significance Johnny Horne, FSU Introduction to Polychaeta: The orders of interest belong to class

Polychaeta, of phylum Annelida. They are a diverse group of metameric worms including over 10,000 species. They are generally marine, schizocoelous, and develop by means of a trocophore larva. The diversity of this class is due in part to the metameric body plan, allowing for specialization of body segments to suit the habits and habitat of the polychaete. Most possess highly developed parapodia on the mid-section or trunk with bristly protrusions called chaetae. Parapodia are modified for crawling, swimming, burrowing, gas exchange, excretion, feeding, and many other features. The coelom is split into the body cavities present in each segment and serves as a hydrostatic skeleton to aid in movement as well as circulation. Most members possess a cerebral ganglion, have a complete digestive tract, and are dioecious. Reference the appendix for an example of the Polychaeta body plan. Reproduction is accomplished asexually as well as sexually by a variety of means, the most peculiar of which is called epitoky(Brusca, 2002). Epitoky is the transformation of an asexual worm into one with reproductive ability, either in part or as a whole, as well as development of several pelagic adaptations in order to carry out reproduction in the water column. This allows benthic polychaetes to reproduce by pelagic means. Epitoky confers an incredible fecundity, and its complex means are at a level of efficiency unseen in creatures of such primitive state(Chatelain. 2000) Methods of Epitoky: Epitoky is observed most commonly in the orders Cirratulida, Eunicida, and Phyllodocida. These taxonomic references are according to those in the text Invertebrates (Brusca, 2002). The species in these orders range an enormous diversity, and their epitokous mechanisms also vary greatly. The two general epitoky strategies used are: 1) Schizogamy- the asexual polychaete produces buds from its posterior segments to function as reproductive individuals and releases them into the water column for spawning. This type of epitoky is iteroparous, meaning it can be repeated several times throughout the life span of the individual and is not necessarily lethal to the parent. This is common among the polychaete order Phyllodocida(Fischer, 1999; Hoffman, 1972). 2) Epigamy- the process by which the entire asexual individual transforms into an epitoke or heteronereid. This type of epitoky is terminal, or semelparous. All somatic processes halt at the onset of epigamy and efforts center on production of the epitokous form and gametogenesis. The epitoke will die upon spawning or shortly after. This strategy is common among the orders Cirratulida and Eunicida(Brusca, 2002; Peterson, 1999). This dramatic display of reproductive power represents an incredible adaptation to the benthic lifestyle. It enables an animal whose existence is almost entirely substrate bound to reproduce in the higher, more mobile levels of the marine habitat at very high rates of success. Schizogamy and epigamy have been subject to intensive study, but

2 epigamy provides several facets conducive to experimentation and therefore has been described more thoroughly in the literature. Shizogamous Epitoky: Schizogamy is often an event synchronized among the entire local population. It can be synchronized by a variety of external parameters such as temperature, marine salinity, and most commonly by lunar periodicity. This phenomenon is observed in the southern Polynesian islands during the months of October, November, and December. For a three day period at the end of the lunar cycle the spawning bodies of Palola viridis will surface within plus or minus 57 minutes of 2:00 am. This mass synchronization illustrates the incredible power of this reproductive capacity. The epitokes all mature and are released at exactly the same time and spawn as a group. The result is an animal whose survival and fecundity is virtually guaranteed(Brusca 2002; Fischer, 1999). A visual of the budding process of this schizogamy is included in the appendix. Schizogamy most often results in several epitokes. Each will be outfitted with pelagic adaptations such as swelled parapodia and enlarged chaetae. Beyond these adaptations many develop a single photoreceptor, which serves to guide the epitoke to the surface for spawning(Fischer, 1999; Hoffman, 1972). Epigamous Epitoky: Epigamy is the delayed final stage of ontogeny followed almost immediately by spawning and death. The parent generation makes the ultimate sacrifice in order to achieve the largest reproductive event possible. This strategy is as complex as it is fascinating, and has captured the attention of many scientists worldwide due to a variety of aspects. Several of these aspects are: 1)The elusive neurosecretory factor 2)Profound shifts in metabolism 3)Histolysis and transdifferentiation of seemingly mature tissues 4)Semelparity (Chatelain, 2000; Fischer, 1999; Golding, 1994; Finch, 1994) 1)The elusive neurosecretory factor- The fine details of the internal onset of epitoky still confounds the scientific community. Although the onset of epitoky can be observed, measured, inhibited, and even prevented the precise nature of its chemical basis, kinetics, and receptor is still unknown. Based on the combination of several studies of endocrine activity, ganglionic transplantation, and biochemical activity it has been concluded that the internal factor is located in the cerebral ganglion and acts to inhibit epitoky until diminished. The endocrine activity levels start at a high level and begin to fade as the animal approaches its epitokous period. When this inhibitory factor is present the animal can undergo regeneration, but in its deficit or absence it loses this ability and the transformations associated with epitoky begin. The implications of this neurosecretion are enormous! This one hormone holds the balance between regeneration and transdifferentiation followed by reproduction and death. The result of isolation of this endocrine factor has great potential for incredible scientific manipulation and experimentation(Chatelain, 2007; Fischer, 1999; Golding, 1994). Unfortunately it has not made a meaningful appearance in the literature for several years.

2)Profound shifts in metabolism- Immediately upon onset of epitoky the metabolism of the animal undergoes a profound shift to prepare for a number of challenges. Somatic processes are abandoned almost entirely. The existing resources are metabolized for use in the transformation and spawning of the epitoke. The newly developed musculature of the epitoke will undergo fervent and high speed contractions during the pelagic stint prior to reproduction. Primarily aerobic means are required to sustain this effort. These conclusions were given evidence by a series of studies continued by Chatelain regarding the biochemistry of atokes and epitokes as well as their mitochondrial variations . The mitochondrial presence was shown to increase by a large factor in both size and number according to metabolic studies done on male epitokes of Nereis virens. Aerobic capacity, measured by change in VO2 max and glutamate dehydrogenase levels in the mitochondria, increased three-fold in epitokous individuals, while the products of anaerobic processes decreased by a factor of seven(Fischer, 1999; Chatelain, 2007). 3)Histolysis and transdifferentiation of seemingly mature tissues-The transformation of epigamous species is marked by a development of pelagic adaptations, as well as histolysis of many existing benthic body forms. During the transdifferentiation the parapodia swell and the chaetae extend- see the appendix for a diagram detailing the parapodial enhancements common to epigamous polychaetes. The musculature of the animal undergoes intensive remodeling. 80% of circular muscle and 30% of longitudinal muscle are diminished entirely to be replaced with more efficient and well developed transverse muscle tissue. The digestive portion of the animal is almost entirely diminished. Feeding and digestion do not take place after the onset of epitoky except in some females who brood the young for a short period during which they must feed. The nervous system undergoes massive expansion in many cases. The result is a more extensive nervous system, and new receptors in many cases. This can be in the form of complex photoreceptors or chemoreceptors. Almost the entirety of the internal portion of the worm is reassigned to the task of gametogenesis(Brusca, 2002; Chatelain 2007; Fischer, 1999; ). 4)Semelparity- an adaptation which emphasizes production of a new generation over survival of the individual. Epigamous polychaetes undergo semelparous epitoky, meaning the parent generation die shortly after spawning in all but a few rare cases. The evolutionary significance of this trait is immense- a benthic invertebrate has adapted a mechanism by which survival of the individual comes second to production of progeny; seemingly clear proof of a feat of large evolutionary fitness. A study was done on the semelparous Nereis diversicolor by Golding and Yuwono in which epitoky was able to be inhibited by implantation of a juvenile cerebral ganglion, and the polychaete would resume asexual somatic activity and re-display regeneration. This resulted in a longevity increase and could be repeated at the next epitokous onset a second and sometimes even a third time. They speculated that this provided evidence of semelparity being a secondary trait and a latent iteroparous ability was present. If this were the case it would show that this primitive invertebrate has been able to adapt reproductive ability capable of multiplying the population at every event of reproduction (Golding, 1994; Finch, 1994; Fischer, 1999).

4 Ecological signifigance of Epitoky: The ecological signifigance of epitoky is evident upon observation of the widespread presence and strength of polychaetes in the marine habitat. By means of this efficient mechanism the population of many orders of polychaetes remains high and their place within the biological realm is seemingly permanent. The large dispersal that is the result of epitoky confers resistance to extinction on a local and global scale. Epitoky is an adaptation mainly adapted by benthic bottom feeders to reproduce within the water column. By packing an epitoke, schizogamous or epigamous, with the nutrients acquired from the substrate a dispersal of the biological wealth is accomplished on a grand scale. The nutrients of the substrate are recycled into the water column and dispersed over a large area. Many pelagic animals rely on these massive spawning events as a food source, directly and indirectly. Evolutionary implications: Epitoky is a unique adaptation and is certainly a hot topic in the area of evolutionary biology. This instance of reproductive fitness allows a benthic animal to produce pelagic reproductive forms. This will increase the area inhabited by the species, in turn lowering chances of extinction events, elminating competition between generations in schizogamous species, and dispersal over a large area while increasing the reproductive success Semelparity is observed in the epigamous species, and the significance of a latent iteroparous capacity seemingly observed by Golding and Yuwono would indicate that this adaptation is a relatively new event in response to a need to increase their population or other dire environmental factors(Golding, 1994; Finch, 1994; Fischer, 1999). Epitoky is an instance of incredibly high fecundity. Synchronized spawning, pelagic developments in the reproductive unit, pelagic dispersal, and devotion of the entirety of the epitoke to gametogenesis results in instances of multiple progeny per parent, and resultant population expansion in number and area is almost immeasurable. The resultant generation is spread over a large area, minimizing extinction possibilities, and in cases of schizogamy will not allow for competition with the parents. Schizogamous species also decrease the chance of attracting predators by releasing the nutritious reproductive buds into the water column. This incident of incredible fitness is evidence of the evolutionary success of these primitive annelids(Chatelain, 2007; Fischer, 1999). Conclusion: In summary, epitoky is an adaptive phenomenon manifested by many polychaete worms which has allowed for their widespread success. This adaptation allows benthic worms to reproduce in the water column by means of a pelagic epitoke, sent as a messenger or in the form of a full body transformation. This climactic process results in a stable and growing population. Evolutionarily epitoky represents an incredible step forward for polychaeta, by which their fecundity has been increased several degrees of magnitude. Ecologically this process is significant because it results in the dispersal of nutrients, biomass, and polychaete worms. Whether it be the sacrificial semelparity of N. Diversicolor or the widespread and predictable spawning of Palola viridis epitoky represents a biological wonder not to be underestimated.

Appendix

The figures above are a representative body plan of polychaeta as well as the visualization of the enhancements parapodia undergo during epitoky. (Brusca, 2002)

To the left is a Palola viridis, undergoing asexual epitoky. The buds are the posterior formations(Brusca, 2002). Works Cited

7 (Brusca, 2002) Brusca and Brusca. Invertebrates. 2002.(Textbook) (Chatelain, 2000) Chatelain. Epitoky in Nereis (Neanthes) virens : A story about sex and death. Comparative biochemistry and ph ysiology. Biochemistry & molecular biology (2000). 2008;149:202208 (Finch, 1994) Finch CE. Latent Capacities for Gametogenic Cycling in the Semelparous Invertebrated Nereis. Proceedings of the National Academy of Sciences - PNAS. 1994;91:11769-11770 (Golding, 1994) Golding D. Latent Capacities for Gametogenic Cycling in the Semelparous Invertebrate Nereis. Proceedings of the National Academy of Sciences - PNAS. 1994;91:11777-11781 (Hoffman, 1972) Hofmann DK. PROOF OF EPITOKY AS MODE OF REPRODUCTION OF POLYCHAETE EUNICE-SICILIENSIS (Polychaeta: Annelida). Marine biology. 1972;14:341-344. (Fischer, 1999) Fischer A. Reproductive and developmental phenomena in annelids: a source of exemplary research problems. Hydrobiologia. 1999;402:1-20. (Peterson, 1999) Petersen ME. Reproduction and development in Cirratulidae (Annelida : Polychaeta). Hydrobiologia. 1999;402:107-128.