2.

Pheromone detection
Pheromones are very diversified molecules, as has been seen in the previous section.
Therefore the recognition system for pheromones needs to be able to detect all the
different types of ligands. This section will elaborate on the mechanisms of pheromone
perception and on the further transduction pathway in the brain.
Pheromones are detected by both the olfactory receptor system and, in mammals,
the vomeronasal organ of Jacobsen (VNO) (Francia et al., 2014). The main olfactory
epithelium (MOE) consists of two types of G-protein coupled receptors (GPCR) and of
Guanylyl cyclase D receptors (Liberles, 2014). The two types that belong to the GPCRs
are the odorant receptor family (OR) and the trace amine-associated receptors (TAARs)
(Mori,1014).These receptors function according to the stereospecific receptor theory.
This theory assumes that odour perception occurs on the condition that the structures of
the receptor and ligand match (Mori, 2014). Next to the olfactory receptor system, most
mammals also possess the vomeronasal organ (VNO). Three receptors have been
identified in the VNO for the recognition of substances: vomeronasal receptor type 1
(V1R), vomeronasal receptor type 2 (V2R) and formyl peptide receptors (FPRs) (Mori,
2014).
2.1 Receptors of the Odorant Receptor family
Firstly, the odorant receptor family is the largest group of receptors present in the
mammalian MOE (Liberles, 2014). It comprises thousands of proteins: these proteins are
able to detect distinctive features in the odorant ligands such as the length, molecular
size and functional groups of the ligand molecules (Mori, 2014). Odorant receptor ligands
are mainly volatile molecules (Liberles, 2014) and it is hypothesized that recognition
mainly occurs by relatively weak hydrophobic interactions (Mori, 2014). Each neuron
expresses one type of receptor (Liberles, 2014). As has been mentioned before, the
odorant receptor is a member of the GPCRs, these consist of a seven-transmembrane
region (Mori, 2014). The activated receptors couple to G-proteins that contain the subunit
G . This activates a cAMP cascade, resulting in membrane depolarization and action
potential generation in the glomeruli of the olfactory bulb (Liberles, 2014).
Secondly, the TAARs function similarly to the odorant receptor family: their GPCR
cascade pathway is identical. However, the amount of TAARs in the MOE is significantly
lower, as is shown in figure 1 (Mori, 2014), and TAARs are more sensitive towards
odorant molecules containing amino groups, such as phenylethylamine and methyl
piperidine. The fact that TAARs in mice are able to detect volatile compounds in urine
can indicate the involvement of TAARs in pheromone recognition (Zufall & LeindersZufall, 2007).
The third type of MOE receptors, Guanylyl cyclase-D receptors, responds to a
different range of stimuli such as CO . CO traverses through the membrane and forms
bicarbonate in the cell. This leads to cGMP synthesis and depolarization, generating
action potentials. Also, Guanylyl cyclase-D responds to extracellular peptides and carbon
-disulfide (Liberles, 2014).
olf

2.2 Receptors of the Vomeronasal Organ

Although humans and mammals both possess the receptors in the MOE, most animals
also possess the vomeronasal organ (VNO) that is specialized for pheromonal detection
(Mori, 2014). It distinguishes itself from the MOE by the structural difference in GPCRs
(Barret et al., 2012). Remarkably, the VNO opens up to both the nasal cavity and the oral
cavity. This indicates that pheromones can be noticed by different manners than solely
by smell (Mori, 2014). The VNO is situated above the palate and it is composed of
cylindrical structures (Liberles, 2014). The epithelium is separated into two layers, an

apical and basal layer. Both layers possess different vomeronasal receptors (Francia et
al., 2014).
The apical layer consists of mainly vomeronasal type 1 receptors (V1R), these
belong to the rhodopsin class A GPCRs. They recognize organic volatile and non-volatile
compounds, including ligands comprised of sulphated steroids such as androgen and
oestrogen (Mori, 2014). V1R utilize G subunits for their cascade pathway and the
molecules phospholipase, carbon, diacylglycerol and the ion channel TrpC2 for
downstream regulation (Liberles, 2014). Their specificity does not widen when stimulus
concentration is increased so, as studies have shown, removal of a number of V1Rs
causes failure to recognize certain pheromonal cues (Zufall & Leinders-Zufall, 2007).
i2

Figure 1. View of the olfactory receptor epithelium and vomeronasal organ. Number represent amount of receptors in
mice. V1R vomeronasal type 1 receptor, V2R vomeronasal type 2 receptor, FPR formyl peptide receptor, OR olfactory
epithelium receptor, TAAR trace amine-associated receptor, GC-D guanylyl cylase D rececptor (Mori, 2014).

The basal epithelium of the VNO is formed by the vomeronasal type 2 receptor
family (V2R). They belong to the class C GPCR family. V2Rs are structurally different
from the V1Rs due to a large N-terminal domain (Mori, 2014) and the fact that they utilize
Go for their cascade pathway (Liberles, 2014). The N-binding site causes V2R to be
sensitive towards larger non-volatile pheromonal cues such as peptides and proteins. A
certain type of peptides to which V2Rs are sensitive are the antigen MHC class 1
peptides. These antigens are connected to the immune system and transduce
information about the genetic information of the transmitter to the receiver. So animals
are able to sense the state of health of the transmitter and genetic similarities between
them both (Zufall & Leinders-Zufall, 2007).

The last type of receptors, the formyl peptide receptors (FPRs), are present in a
significantly lower number in the VNO than the other types of receptors. These receptors
are specialized in the recognition of infected microorganisms and diseases (Mori, 2014).
However, studies conducting ligand binding analyses have not shown matching results.
They agree that FPRs detect bacterial peptides but their research disagreed on the type
of activating peptides. The function of FPRs is still relatively elusive (Liberles, 2014).
2.3 Allohormone reception
Although the vomeronasal organ and the olfactory system are the main receivers for
pheromones, there is also another way of perceiving them. As has been mentioned
before, these are allohormone pheromones. These pheromones are deposited directly
on the tissue of the receiver, thereby not using the chemosensory cells. The exact
receptor targets of the protein allohormones is not exactly known, only that they can be
delivered to the bloodstream by scratches in certain salamander species (Wyatt, 2014).
2.4 Further procession of pheromones within the neural circuit
Input from the odorant receptor family causes depolarization of second order neurons,
called mitral and tufted cells, in the spherically structured glomeruli of the main olfactory
bulb. In mice there are more than 2000 glomeruli present and they receive about 5000
axons of sensory neurons. TAARs and GC-D target different places in the main olfactory
bulb. However, their connection with other brain structures remains unknown.
Input from the VNO is processed in the glomerular layer of the adjacent olfactory
bulb. The V1R and V2R project into separate sites in the adjacent bulb. A part of these
signals is then further processed in the higher limbic systems.
The mitral cells from both the adjacent and main olfactory bulb project deeper into
the brain. The cells of the MOB innervate the piriform cortex, cortical amygdala and
olfactory tubercle. The adjacent olfactory bulb processes its signals towards the
vomeronasal amygdala, positioned in the limbic brain structures. Sex or state-dependent
responses are regulated in the limbic systems, this occurs because the inputs from the
VNO are further hormonally regulated. The VNO pheromones evoke two types of
responses in animals: endocrine and behavioural responses. Examples of endocrine
responses are the secretion of hypothalamic neurons such as LH, dopamine and
glucocorticoids. One example of endocrine responses is the onset of hormonal maturity
in animals: such an effect is the Whittenberg effect. This causes urinary secretions from
males to synchronize oestrous development in a group of females. Also, experiments
have shown that puberty in females can be accelerated upon encounters with male
odours; this is called the Vandenbergh effect. Furthermore behavioural responses
include sex drive, aggression and parental responsibility (Liberles, 2014).
Currently, researchers agree upon the perception of pheromones via the olfactory
epithelium and the VNO in animals, however, the perception of pheromones in humans
is still subject to debate. This will be further discussed in the upcoming sections.