Peter A. Brennan

Something in the Air? New Insights into Mammalian Pheromones

Olfaction is the dominant sensory modality for most animals and chemosensory communication is particularly well developed in many mammals. Our understanding of this form of communication has grown rapidly over the last ten years since the identification of the first olfactory receptor genes. The subsequent cloning of genes for rodent vomeronasal receptors, which are important in pheromone detection, has revealed an unexpected diversity of around 250 receptors belonging to two structurally different classes. This review will focus on the chemical nature of mammalian pheromones and the complementary roles of the main olfactory system and vomeronasal system in mediating pheromonal responses. Recent studies using genetically modified mice and electrophysiological recordings have highlighted the complexities of chemosensory communication via the vomeronasal system and the role of this system in handling information about sex and genetic identity. Although the vomeronasal organ is often regarded as only a pheromone detector, evidence is emerging that suggests it might respond to a much broader variety of chemosignals.

The nose knows who's who: chemosensory individuality and mate recognition in mice

Individual recognition is an important component of behaviors, such as mate choice and maternal bonding that are vital for reproductive success. This article highlights recent developments in our understanding of the chemosensory cues and the neural pathways involved in individuality discrimination in rodents. There appear to be several types of chemosensory signal of individuality that are influenced by the highly polymorphic families of major histocompatibility complex (MHC) proteins or major urinary proteins (MUPs). Both have the capability of binding small molecules and may influence the individual profile of these chemosignals in biological fluids such as urine, skin secretions, or saliva. Moreover, these proteins, or peptides associated with them, can be taken up into the vomeronasal organ (VNO) where they can potentially interact directly with the vomeronasal receptors. This is particularly interesting given the expression of major histocompatibility complex Ib proteins by the V2R class of vomeronasal receptor and the highly selective responses of accessory olfactory bulb (AOB) mitral cells to strain identity. These findings are consistent with the role of the vomeronasal system in mediating individual discrimination that allows mate recognition in the context of the pregnancy block effect. This is hypothesized to involve a selective increase in the inhibitory control of mitral cells in the accessory olfactory bulb at the first level of processing of the vomeronasal stimulus.

Low molecular weight constituents of male mouse urine mediate the pregnancy block effect and convey information about the identity of the mating male

Mouse urine contains a complex mixture of chemosignals including a variety of small volatile molecules that are bound to major urinary proteins. In addition to signalling maleness, male urine also conveys information about individuality, which allows recently mated female mice to distinguish the urinary chemosignals of the mating male from those of an unfamiliar male. The highly polymorphic nature of the major urinary proteins makes them a likely candidate for conveying individuality information in the context of the pregnancy block effect. This was investigated by comparing the pregnancy‐blocking effectiveness of a high molecular weight urinary fraction, containing major urinary proteins, with that of a low molecular weight fraction containing volatile ligands. Not only was the high molecular weight fraction ineffective in blocking pregnancy, but it also appeared to be less important in signalling individuality than the low molecular fraction. The high molecular weight fraction was ineffective in inducing expression of the immediate early gene product egr‐1 in the accessory olfactory bulb. In contrast, the low molecular weight fraction induced egr‐1 expression in the mitral/tufted neurons in the anterior subregion of the accessory olfactory bulb, suggesting that they activate the V1R class of vomeronasal receptor neuron.

Selective ablation of olfactory receptor neurons without functional impairment of vomeronasal receptor neurons in OMP-ntr transgenic mice

This study used transgenic mice, in which expression of a bacterial nitroreductase (ntr) gene was linked to the expression of olfactory marker protein (OMP). The nitroreductase enzyme is thus expressed in mature chemosensory neurons of these OMP‐ntr transgenic mice, and converts the pro‐drug CB1954 to a cytotoxic form, specifically killing these neurons. Systemic injections of the pro‐drug led to the ablation of receptor neurons in both the main olfactory and vomeronasal epithelia. Due to the anatomical separation of the epithelia, however, when the pro‐drug was administered by intranasal infusion only the receptors of the main olfactory epithelium were destroyed. This procedure resulted in a profound deficit in olfactory investigation and discrimination in a habituation–dishabituation test, whereas the pregnancy blocking effect of male pheromones, which is mediated via the vomeronasal system was unaffected. OMP‐ntr mice receiving intranasal infusion of pro‐drug had not recovered any significant main olfactory function at 24 days following treatment. This novel technique could potentially be applied to selectively ablate olfactory receptor neurons expressing a particular olfactory receptor by linking its expression to that of the nitroreductase enzyme.

Changes in electrophysiological activity in the accessory olfactory bulb and medial amygdala associated with mate recognition in mice

The ability of female mice to recognize their mates pheromonal identity is critical for the maintenance of their pregnancy and is hypothesized to involve increases in the inhibitory control of mitral/tufted projection neurons in the accessory olfactory bulb. Local field potential recordings from this region of freely behaving female mice showed oscillating neural activity over a wide range of frequencies, which was affected by chemosensory input and prior experience. Mating caused lasting increases in the baseline neural activity in the accessory olfactory bulb, with large increases in the amplitude of local field potential oscillations across a range of frequencies. Exposure to the mates urinary cues remained effective in increasing the power of these oscillations following mating, but urinary cues from an unfamiliar male were ineffective. A differential response to the familiar and unfamiliar chemosignals was also observed at the level of the amygdala following mating. Individual neurons in the medial amygdala responded more strongly to urine from an unfamiliar male than from the mating male. These findings are consistent with the selective enhancement of inhibition of the familiar pheromonal signal at the level of the accessory olfactory bulb, which is proposed to underlie recognition of the mating male.

Retrograde labelling of mitral/tufted cells in the mouse accessory olfactory bulb following local injections of the lipophillic tracer DiI into the vomeronasal amygdala☆

It has recently become apparent that there are two classes of vomeronasal receptor neurons that project to functionally separate anterior and posterior sub-regions of the mammalian accessory olfactory bulb. However, anterograde tracing of the projections from these sub-regions, in the mouse, has revealed that the processing pathways are not segregated at the level of the vomeronasal amygdala. Both sub-regions have overlapping projections to the superficial lamina of the medial and posterior medial cortical nuclei of the amygdala. However, differential projections have been found in the opossum, in which only the posterior sub-region projects to the deeper laminae of the medial amygdala. Therefore, there may be species differences in these projections that are important for the control of reproductive behaviour. This study used an alternative approach of retrogradely tracing mitral/tufted cell projections from different nuclei of the vomeronasal amygdala back to the accessory olfactory bulb of mice. Local injections of the lipophilic tracer DiI were made into the antero-dorsal and postero-ventral divisions of the medial amygdala, and into the postero-medial cortical amygdala. In each case, provided the DiI affected the superficial lamina Ia, labelled mitral/tufted cells were found distributed throughout the anterior-posterior extent of the accessory olfactory bulb. These results confirm that mitral/tufted cells of the anterior and posterior sub-regions of the accessory olfactory bulb project to both the medial and postero-medial cortical nuclei of the amygdala. There was no evidence for differential projections from the anterior and posterior sub-regions accessory olfactory bulb in mice, as has been reported to occur in other species.

Animal behaviour: How mice make their mark

The social behaviour of many animals relies on their ability to use odour cues to distinguish among individuals. Studies of mice highlight the importance of urinary proteins in this complex signalling system.