Kevin R. Kelliher

Essential Role of the Main Olfactory System in Social Recognition of Major Histocompatibility Complex Peptide Ligands

Genes of the major histocompatibility complex (MHC), which play a critical role in immune recognition, influence mating preference and other social behaviors in fish, mice, and humans via chemical signals. The cellular and molecular mechanisms by which this occurs and the nature of these chemosignals remain unclear. In contrast to the widely held view that olfactory sensory neurons (OSNs) in the main olfactory epithelium (MOE) are stimulated by volatile chemosignals only, we show here that nonvolatile immune system molecules function as olfactory cues in the mammalian MOE. Using mice with targeted deletions in selected signal transduction genes (CNGA2, CNGA4), we used a combination of dye tracing, electrophysiological, Ca2+ imaging, and behavioral approaches to demonstrate that nonvolatile MHC class I peptides activate subsets of OSNs at subnanomolar concentrations in vitro and affect social preference of male mice in vivo. Both effects depend on the cyclic nucleotide-gated (CNG) channel gene CNGA2, the function of which in the nose is unique to the main population of OSNs. Disruption of the modulatory CNGA4 channel subunit reveals a profound defect in adaptation of peptide-evoked potentials in the MOE. Because sensory neurons in the vomeronasal organ (VNO) also respond to MHC peptides but do not express CNGA2, distinct mechanisms are used by the mammalian main and accessory olfactory systems for the detection of MHC peptide ligands. These results suggest a general role for MHC peptides in chemical communication even in those vertebrates that lack a functional VNO.

Social motivation is reduced in vasopressin 1b receptor null mice despite normal performance in an olfactory discrimination task

In this study, we characterized more thoroughly the social behavior of vasopressin 1b receptor null (V1bR-/-) mice. We confirmed that V1bR-/- males exhibit less social aggression than their wild-type (V1bR+/+) littermates. We tested social preference by giving male subjects a choice between pairs of soiled or clean bedding. In general, V1bR+/+ mice spent significantly more time engaged in chemoinvestigation of these social stimuli than V1bR-/- mice. Male V1bR+/+ mice preferred female-soiled bedding over male-soiled bedding, male-soiled bedding over clean bedding, and female-soiled bedding over clean bedding. In contrast, V1bR-/- males failed to exhibit a preference for any bedding. This difference in behavior is not explained by an anosmic condition as there were no differences between V1bR-/- and V1bR+/+ mice in their abilities to detect a cookie buried in clean bedding, or in their ability to perform in an operant conditioning task using a fully automated liquid dilution olfactometer. In the latter task, male V1bR-/- mice were fully capable of discriminating between male and female mouse urine. The latencies to learn this task did not differ between the two genotypes. Thus, a V1bR-/- males ability to differentiate between male and female chemosensory cues appears no different than that of a V1bR+/+ males. We propose that the V1bR plays an important role in social motivation, perhaps by coupling the processing, integration, and/or interpretation of chemosensory cues with the appropriate behavioral response.

Complementary Roles of the Main and Accessory Olfactory Systems in Mammalian Mate Recognition

We review studies conducted in mouse and ferret that have specified roles of both the main and the accessory olfactory nervous systems in the detection and processing of body odorants (e.g., urinary pheromones, extraorbital lacrimal gland secretions, major histocompatibility complex peptide ligands, and anal scent gland secretions) that play an essential role in sex discrimination and attraction between males and females leading to mate choice and successful reproduction. We also review literature that compares the forebrain processing of inputs from the two olfactory systems in the two sexes that underlies heterosexual partner preferences. Finally, we review experiments that raise the possibility that body odorants detected by the main olfactory system contribute to mate recognition in humans.

Pheromonal recognition memory induced by TRPC2‐independent vomeronasal sensing

Detection and transduction of pheromonal signals by the mouse vomeronasal organ (VNO) is critical for the formation of a persistent memory required for mate recognition in the context of selective pregnancy failure (the Bruce effect). This pregnancy block can be mediated by peptide ligands of disparate major histocompatibility complex (MHC) molecules, but little is known about the molecular mechanisms underlying this effect. Given the proposed key role of the transient receptor potential channel, TRPC2, in VNO signal transduction, we tested whether TRPC2 is essential for memory formation in the context of the Bruce effect. Surprisingly, the loss of the TRPC2 channel did not significantly influence memory formation, whereas surgical lesions of the VNO caused a profound deficit. Furthermore, field potential and single‐cell patch‐clamp recordings showed that TRPC2 is dispensable for the transduction of MHC peptide ligands by sensory neurons in the basal zone of the VNO. This indicates that a previously unrecognized TRPC2‐independent signal transduction mechanism in the VNO underlies the sensing of cues required for the formation of this pheromonal recognition memory.

An olfactory subsystem that detects carbon disulfide and mediates food-related social learning

Olfactory signals influence social interactions in a variety of species. In mammals, pheromones and other social cues can promote mating or aggression behaviors; can communicate information about social hierarchies, genetic identity and health status; and can contribute to associative learning. However, the molecular, cellular, and neural mechanisms underlying many olfactory-mediated social interactions remain poorly understood. Here, we report that a specialized olfactory subsystem that includes olfactory sensory neurons (OSNs) expressing the receptor guanylyl cyclase GC-D, the cyclic nucleotide-gated channel subunit CNGA3, and the carbonic anhydrase isoform CAII (GC-D(+) OSNs) is required for the acquisition of socially transmitted food preferences (STFPs) in mice. Using electrophysiological recordings from gene-targeted mice, we show that GC-D(+) OSNs are highly sensitive to the volatile semiochemical carbon disulfide (CS(2)), a component of rodent breath and a known social signal mediating the acquisition of STFPs. Olfactory responses to CS(2) are drastically reduced in mice lacking GC-D, CNGA3, or CAII. Disruption of this sensory transduction cascade also results in a failure to acquire STFPs from either live or surrogate demonstrator mice or to exhibit hippocampal correlates of STFP retrieval. Our findings indicate that GC-D(+) OSNs detect chemosignals that facilitate food-related social interactions.

Parallel processing of social signals by the mammalian main and accessory olfactory systems

Abstract.The mammalian olfactory system has evolved complex mechanisms to detect a vast range of molecular cues. In rodents, the olfactory system comprises several distinct subsystems. Current interest has focused on the exact role that each of these subsystems plays in detecting molecular information and regulating chemosensorydependent behaviors. Here, we summarize recent results showing that the mouse main and accessory olfactory systems detect, at least in part, overlapping sets of social chemosignals. These findings give rise to a model that involves parallel processing of the same molecular cues in both systems. Together with previous work, this model will lead to a better understanding of the general organization of chemical communication in mammals and give a new direction for future experiments.

Signaling in the Chemosensory Systems

Abstract.The mammalian olfactory system has evolved complex mechanisms to detect a vast range of molecular cues. In rodents, the olfactory system comprises several distinct subsystems. Current interest has focused on the exact role that each of these subsystems plays in detecting molecular information and regulating chemosensorydependent behaviors. Here, we summarize recent results showing that the mouse main and accessory olfactory systems detect, at least in part, overlapping sets of social chemosignals. These findings give rise to a model that involves parallel processing of the same molecular cues in both systems. Together with previous work, this model will lead to a better understanding of the general organization of chemical communication in mammals and give a new direction for future experiments.

Importance of the CNGA4 channel gene for odor discrimination and adaptation in behaving mice

Odor stimulation of olfactory sensory neurons (OSNs) leads to both the activation and subsequent desensitization of a heteromultimeric cyclic-nucleotide-gated (CNG) channel present in these cells. The native olfactory CNG channel consists of three distinct subunits: CNGA2, CNGA4, and CNGB1b. Mice in which the CNGA4 gene has been deleted display defective Ca2+/calmodulin-dependent inhibition of the CNG channel, resulting in a striking reduction in adaptation of the odor-induced electrophysiological response in the OSNs. These mutants therefore afford an excellent opportunity to assess the importance of Ca2+-mediated CNG channel desensitization for odor discrimination and adaptation in behaving animals. By using an operant conditioning paradigm, we show that CNGA4-null mice are profoundly impaired in the detection and discrimination of olfactory stimuli in the presence of an adapting background odor. The extent of this impairment depends on both the concentration and the molecular identity of the adapting stimulus. Thus, Ca2+-dependent desensitization of the odor response in the OSNs mediated by the CNGA4 subunit is essential for normal odor sensation and adaptation of freely behaving mice, preventing saturation of the olfactory signal transduction machinery and extending the range of odor detection and discrimination.

The combined role of the main olfactory and vomeronasal systems in social communication in mammals.

The main olfactory and the vomeronasal systems are the two systems by which most vertebrates detect chemosensory cues that mediate social behavior. Much research has focused on how one system or the other is critical for particular behaviors. This has lead to a vision of two distinct and complexly autonomous olfactory systems. A closer look at research over the past 30 years reveals a different picture however. These two seemingly distinct systems are much more integrated than previously thought. One novel set of chemosensory cues in particular (MHC Class I peptide ligands) can show us how both systems are capable of detecting the same chemosensory cues, through different mechanisms yet provide the same general information (genetic individuality). Future research will need to now focus on how two seemingly distinct chemosensory systems together detect pheromones and mediate social behaviors. Do these systems work independently, synergistically or competitively in communicating between individuals of the same species?

The ferret's vomeronasal organ and accessory olfactory bulb: effect of hormone manipulation in adult males and females.

The male ferret, a carnivore, was recently shown to possess a vomeronasal organ (VNO). We compared the morphology of the VNO and its associated accessory olfactory bulb (AOB) in male and female ferrets that were killed in adulthood. The volume and surface area of the VNO neuroepithelium were similar in adult gonadectomized male and female ferrets regardless of whether they were treated with testosterone propionate (TP) or oil vehicle. An AOB was localized bilaterally in the medial caudal part of the olfactory bulbs of adult ferrets using soybean agglutin binding and immunostaining for luteinizing hormone‐releasing hormone and tyrosine hydroxylase as well as Nissl staining of coronal, horizontal, and sagittal brain sections. There was no effect of sex or TP treatment on AOB cell layer volume in adult gonadectomized animals. We found the ferrets AOB to be more medially located and much smaller than previously reported in this species, thus highlighting the importance of using several histochemical markers to characterize this structure in any previously unexamined species. Adult male and female ferrets both have a VNO and an associated AOB. More research is needed to determine what role, if any, this accessory olfactory system plays in mediating behavioral and neuroendocrine responses to pheromones in ferrets of either sex. Anat Rec 263:280–288, 2001.