Vomeronasal Information Research Articles

Neuroanatomical and Immunohistological Study of the Main and Accessory Olfactory Bulbs of the Meerkat (Suricata suricatta).

Simple SummaryIn wild mammals, chemical senses are crucial to survival, but sensory system information is lacking for many species, including the meerkat (Suricata suricatta), an iconic mammal with a marked social hierarchy that has been ambiguously classified in both canid and felid families. We studied the neuroanatomical basis of the meerkat olfactory and accessory olfactory bulbs, aiming to provide information on the relevance of both systems to the behaviors of this species and contributing to improving its taxonomic classification. The accessory olfactory bulb serves as the integration center of vomeronasal information. When examined microscopically, the accessory olfactory bulb of the meerkat presents a lamination pattern more defined than observed in dogs and approaching the pattern described in cats. The degree of lamination and development in the meerkat main olfactory bulb is comparable to the general pattern observed in mammals but with numerous specific features. Our study supports the functionality of the olfactory and vomeronasal integrative centers in meerkats and places this species within the suborder Feliformia. Our study also confirms the importance of chemical signals in mediating the social behaviors of this species and provides essential neuroanatomical information for understanding the functioning of their chemical senses.We approached the study of the main (MOB) and accessory olfactory bulbs (AOB) of the meerkat (Suricata suricatta) aiming to fill important gaps in knowledge regarding the neuroanatomical basis of olfactory and pheromonal signal processing in this iconic species. Microdissection techniques were used to extract the olfactory bulbs. The samples were subjected to hematoxylin-eosin and Nissl stains, histochemical (Ulex europaeus agglutinin, Lycopersicon esculentum agglutinin) and immunohistochemical labelling (Gαo, Gαi2, calretinin, calbindin, olfactory marker protein, glial fibrillary acidic protein, microtubule-associated protein 2, SMI-32, growth-associated protein 43). Microscopically, the meerkat AOB lamination pattern is more defined than the dog’s, approaching that described in cats, with well-defined glomeruli and a wide mitral-plexiform layer, with scattered main cells and granular cells organized in clusters. The degree of lamination and development of the meerkat MOB suggests a macrosmatic mammalian species. Calcium-binding proteins allow for the discrimination of atypical glomerular subpopulations in the olfactory limbus between the MOB and AOB. Our observations support AOB functionality in the meerkat, indicating chemosensory specialization for the detection of pheromones, as identified by the characterization of the V1R vomeronasal receptor family and the apparent deterioration of the V2R receptor family.

Does a third intermediate model for the vomeronasal processing of information exist? Insights from the macropodid neuroanatomy

The study of the α-subunit of Gi2 and Go proteins in the accessory olfactory bulb (AOB) was crucial for the identification of the two main families of vomeronasal receptors, V1R and V2R. Both families are expressed in the rodent and lagomorph AOBs, according to a segregated model characterized by topographical anteroposterior zonation. Many mammal species have suffered from the deterioration of the Gαo pathway and are categorized as belonging to the uniform model. This scenario has been complicated by characterization of the AOB in the tammar wallaby, Notamacropus eugenii, which appears to follow a third model of vomeronasal organization featuring exclusive Gαo protein expression, referred to as the intermediate model, which has not yet been replicated in any other species. Our morphofunctional study of the vomeronasal system (VNS) in Bennett’s wallaby, Notamacropus rufogriseus, provides further information regarding this third model of vomeronasal transduction. A comprehensive histological, lectin, and immunohistochemical study of the Bennett’s wallaby VNS was performed. Anti-Gαo and anti-Gαi2 antibodies were particularly useful because they labeled the transduction cascade of V2R and V1R receptors, respectively. Both G proteins showed canonical immunohistochemical labeling in the vomeronasal organ and the AOB, consistent with the anterior–posterior zonation of the segregated model. The lectin Ulex europaeus agglutinin selectively labeled the anterior AOB, providing additional evidence for the segregation of vomeronasal information in the wallaby. Overall, the VNS of the Bennett’s wallaby shows a degree of differentiation and histochemical and neurochemical diversity comparable to species with greater VNS development. The existence of the third intermediate type in vomeronasal information processing reported in Notamacropus eugenii is not supported by our lectin-histochemical and immunohistochemical findings in Notamacropus rufogriseus.

Integrating pheromonal and spatial information in the amygdalo-hippocampal network

Vomeronasal information is critical in mice for territorial behavior. Consequently, learning the territorial spatial structure should incorporate the vomeronasal signals indicating individual identity into the hippocampal cognitive map. In this work we show in mice that navigating a virtual environment induces synchronic activity, with causality in both directionalities, between the vomeronasal amygdala and the dorsal CA1 of the hippocampus in the theta frequency range. The detection of urine stimuli induces synaptic plasticity in the vomeronasal pathway and the dorsal hippocampus, even in animals with experimentally induced anosmia. In the dorsal hippocampus, this plasticity is associated with the overexpression of pAKT and pGSK3β. An amygdalo-entorhino-hippocampal circuit likely underlies this effect of pheromonal information on hippocampal learning. This circuit likely constitutes the neural substrate of territorial behavior in mice, and it allows the integration of social and spatial information.

Dendritic Organization of Olfactory Inputs to Medial Amygdala Neurons.

The medial amygdala (MeA) is a central hub in the olfactory neural network. It receives vomeronasal information directly from the accessory olfactory bulb (AOB) and main olfactory information largely via odor-processing regions such as the olfactory cortical amygdala (CoA). How these inputs are processed by MeA neurons is poorly understood. Using the GAD67-GFP mouse, we show that MeA principal neurons receive convergent AOB and CoA inputs. Somatically recorded AOB synaptic inputs had slower kinetics than CoA inputs, suggesting that they are electrotonically more distant. Field potential recording, pharmacological manipulation, and Ca(2+) imaging revealed that AOB synapses are confined to distal dendrites and segregated from the proximally located CoA synapses. Moreover, unsynchronized AOB inputs had significantly broader temporal summation that was dependent on the activation of NMDA receptors. These findings show that MeA principal neurons process main and accessory olfactory inputs differentially in distinct dendritic compartments. Significance statement: In most vertebrates, olfactory cues are processed by two largely segregated neural pathways, the main and accessory olfactory systems, which are specialized to detect odors and nonvolatile chemosignals, respectively. Information from these two pathways ultimately converges at higher brain regions, one of the major hubs being the medial amygdala. Little is known about how olfactory inputs are processed by medial amygdala neurons. This study shows that individual principal neurons in this region receive input from both pathways and that these synapses are spatially segregated on their dendritic tree. We provide evidence suggesting that this dendritic segregation leads to distinct input integration and impact on neuronal output; hence, dendritic mechanisms control olfactory processing in the amygdala.

Sex-specific processing of social cues in the medial amygdala.

Animal-animal recognition within, and across species, is essential for predator avoidance and social interactions. Despite its essential role in orchestrating responses to animal cues, basic principles of information processing by the vomeronasal system are still unknown. The medial amygdala (MeA) occupies a central position in the vomeronasal pathway, upstream of hypothalamic centers dedicated to defensive and social responses. We have characterized sensory responses in the mouse MeA and uncovered emergent properties that shed new light onto the transformation of vomeronasal information into sex- and species-specific responses. In particular, we show that the MeA displays a degree of stimulus selectivity and a striking sexually dimorphic sensory representation that are not observed in the upstream relay of the accessory olfactory bulb (AOB). Furthermore, our results demonstrate that the development of sexually dimorphic circuits in the MeA requires steroid signaling near the time of puberty to organize the functional representation of sensory stimuli.DOI: http://dx.doi.org/10.7554/eLife.02743.001.

The vomeronasal cortex – afferent and efferent projections of the posteromedial cortical nucleus of the amygdala in mice

Most mammals possess a vomeronasal system that detects predominantly chemical signals of biological relevance. Vomeronasal information is relayed to the accessory olfactory bulb (AOB), whose unique cortical target is the posteromedial cortical nucleus of the amygdala. This cortical structure should therefore be considered the primary vomeronasal cortex. In the present work, we describe the afferent and efferent connections of the posteromedial cortical nucleus of the amygdala in female mice, using anterograde (biotinylated dextranamines) and retrograde (Fluorogold) tracers, and zinc selenite as a tracer specific for zinc-enriched (putative glutamatergic) projections. The results show that the posteromedial cortical nucleus of the amygdala is strongly interconnected not only with the rest of the vomeronasal system (AOB and its target structures in the amygdala), but also with the olfactory system (piriform cortex, olfactory-recipient nuclei of the amygdala and entorhinal cortex). Therefore, the posteromedial cortical nucleus of the amygdala probably integrates olfactory and vomeronasal information. In addition, the posteromedial cortical nucleus of the amygdala shows moderate interconnections with the associative (basomedial) amygdala and with the ventral hippocampus, which may be involved in emotional and spatial learning (respectively) induced by chemical signals. Finally, the posteromedial cortical nucleus of the amygdala gives rise to zinc-enriched projections to the ventrolateral septum and the ventromedial striatum (including the medial islands of Calleja). This pattern of intracortical connections (with the olfactory cortex and hippocampus, mainly) and cortico-striatal excitatory projections (with the olfactory tubercle and septum) is consistent with its proposed nature as the primary vomeronasal cortex.

Molecular organization of vomeronasal chemoreception.

The vomeronasal organ (VNO) plays a key role in mediating the social and defensive responses of many terrestrial vertebrates to species- and sex-specific chemosignals1. Over 250 putative pheromone receptors have been identified in the mouse VNO2,3, but the nature of the signals detected by individual VNO receptors has not yet been elucidated. In order to gain insight into the molecular logic of VNO detection leading to mating, aggression, or defensive responses, we sought to uncover the response profiles of individual vomeronasal receptors to a wide range of animal cues. We describe here the repertoire of ethological and physiological stimuli detected by a large number of individual vomeronasal receptors, and define a global map of vomeronasal signal detection. We demonstrate that the two classes of vomeronasal receptors V1Rs and V2Rs use fundamentally different strategies to encode chemosensory information, and that distinct receptor subfamilies have evolved towards the specific recognition of certain animal groups or chemical structures. The association of large subsets of vomeronasal receptors with cognate, ethologically and physiologically relevant stimuli establishes the molecular foundation of vomeronasal information coding, and opens new avenues for further investigating the neural mechanisms underlying behavior specificity.

Lungfishes, like tetrapods, possess a vomeronasal system

The vomeronasal system (VNS) is an accessory olfactory system that in tetrapod vertebrates is composed of specific receptor neurons in the nasal organ and a set of centers in the forebrain that receive and relay the information consecutively towards the hypothalamus. Thus, only in tetrapods the VNS comprises a discrete vomeronasal (Jacobson's) organ, which contains receptor cells that are morphologically distinct from those of the olfactory epithelium and use different transduction mechanisms. The axons of the vomeronasal receptors in tetrapods project to the accessory olfactory bulb (AOB) in the rostral telencephalon. Secondary vomeronasal connections exist through the medial amygdala to the hypothalamus. Currently, the lungfishes are considered the closest living relatives of tetrapods. Here we show that the African lungfish, Protopterus dolloi, has epithelial crypts at the base of the lamellae of the olfactory epithelium that express markers of the vomeronasal receptors in tetrapods. The projections of these crypts allow us to identify an AOB on the lateral margin of the main olfactory bulb. The projections of this AOB reach a region that is topologically, hodologically, and immunohistochemically identical to the medial amygdala and could represent its homolog. Neurons of this putative medial amygdala were demonstrated to project to the lateral hypothalamus, as they do in tetrapods. All these features that lungfishes share with tetrapods indicate that lungfishes have the complete set of brain centers and connections involved in processing vomeronasal information and that these features were already present in the last common ancestor of lungfishes and tetrapods.

Lateral and medial amygdala of anuran amphibians and their relation to olfactory and vomeronasal information

The amygdala of anurans is currently considered as a complex of nuclei that share many features with their counterparts in amniotes. In the present study, the subdivisions of the amygdala that are directly related to olfactory and vomeronasal information, were investigated in the anurans Rana perezi and Xenopus laevis. In particular, the connectivity of the main and accessory olfactory bulbs and their related amygdaloid nuclei was studied by means of in vivo and in vitro tract-tracing with dextran amines. The projections observed from the main olfactory bulb clearly innervate the newly redefined lateral amygdala within the ventral pallium and, to a lesser extent, the rostral portion of the medial amygdala. Injections into the accessory olfactory bulb exclusively revealed projections to the medial amygdala. Tracer applications into the lateral and medial nuclei revealed abundant intra-amygdaloid connections. The dual flow of olfactory and vomeronasal projections throughout the telencephalon was not strictly segregated since the lateral pallium and the lateral amygdala, both receiving olfactory information, were found to project to the medial amygdala (the only target of vomeronasal information), which in turn projects to the lateral amygdala. Additionally, both the lateral and the medial amygdala strongly project to the hypothalamus through the anuran equivalent of the stria terminalis. The main hodological features found in the present study suggest that forerunners of the olfactory and vomeronasal amygdaloid nuclei can be distinguished in anurans. This supports the notion that all tetrapods share a common pattern of organization of the amygdaloid complex, which links environmental (olfactory/vomeronasal) information and the behavioural response of the animal.

The “olfactostriatum” of snakes: A basal ganglia vomeronasal structure in tetrapods

The olfactostriatum is a portion of the basal ganglia of snakes situated ventromedially to the nucleus accumbens proper. It receives a major vomeronasal input from the nucleus sphericus, the primary target of accessory olfactory bulb efferents. Recently, the ophidian olfactostriatum has been characterized on the basis of chemoarchitecture (distribution of serotonin, neuropeptide Y and tyrosine hydroxylase) and hodology (afferent and efferent connections). In contrast to the nucleus accumbens proper, the olfactostriatum is densely immunoreactive for serotonin and neuropeptide Y and sparsely immunoreactive for tyrosine hydroxylase. The nucleus accumbens proper and the olfactostriatum share most afferent connections except those originating in the nucleus sphericus, which are exclusively directed to the olfactostriatum. Similarly, the nucleus accumbens proper and the olfactostriatum show a similar pattern of efferent connections including those going to the ventral pallidum, although the olfactostriatum alone projects to the main and accessory olfactory bulbs as well as some amygdaloid nuclei. On the basis of its chemoarchitecture, the olfactostriatum resembles the mammalian ventral pallidum (but also the shell of the nucleus accumbens). Its connections, however, suggests that the olfactostriatum could be a specialized portion of the shell of nucleus accumbens extended more ventromedially than previously believed and devoted to processing vomeronasal information. Comparative data suggest that a similar structure is present in the basal ganglia of amphibians and mammals.

Chemoarchitecture and afferent connections of the “olfactostriatum”: a specialized vomeronasal structure within the basal ganglia of snakes

The olfactostriatum, a portion of the striatal complex of snakes, is the major tertiary vomeronasal structure in the ophidian brain, receiving substantial afferents from the nucleus sphericus, the primary target of accessory olfactory bulb efferents. In the present study, we have characterized the olfactostriatum of garter snakes ( Thamnophis sirtalis) on the basis of chemoarchitecture (distribution of serotonin, neuropeptide Y and tyrosine hydroxylase) and hodology (afferent connections). The olfactostriatum is densely immunoreactive for serotonin and neuropeptide Y and shows moderate-to-weak immunoreactivity for tyrosine hydroxylase. In addition to afferents from the nucleus sphericus, the olfactostriatum receives inputs from the dorsal and lateral cortices, nucleus of the accessory olfactory tract, external and dorsolateral amygdalae, dorsomedial thalamic nucleus, ventral tegmental area and raphe nuclei. Double labeling experiments demonstrated that the distribution of serotonin and neuropeptide Y in this area almost completely overlaps the terminal field of projections from the nucleus sphericus. Also, serotonergic and dopaminergic innervation of the olfactostriatum likely arise, respectively, from the raphe nuclei and the ventral tegmental area, whereas local circuit neurons originate the neuropeptide Y immunoreactivity. These results indicate that the olfactostriatum of snakes could be a portion of the nucleus accumbens, with features characteristic of the accumbens shell, devoted to processing vomeronasal information. Comparative data suggest that a similar structure is present in the ventral striatum of amphibians and mammals.

Neural substrates for processing chemosensory information in snakes

Snakes interact with their chemical environment through their olfactory and vomeronasal systems. The present report summarizes advances on neural substrates for processing chemosensory information. First, the efferent and centrifugal afferent connections of the main and accessory olfactory bulbs were reinvestigated. Second, the afferent and efferent connections of the nucleus sphericus, the main target of the accessory olfactory bulb, were characterized. The nucleus sphericus gives rise to a very small projection to the hypothalamus, but it does project to other telencephalic structures where olfactory and vomeronasal information could converge. Third, the intra-amygdaloid circuitry and the amygdalo-hypothalamic projections were described. The medial amygdala, for instance, receives both vomeronasal and olfactory inputs and projects to the hypothalamus, namely, to the lateral posterior hypothalamic nucleus. Fourth, because the lateral posterior hypothalamic nucleus projects to the hypoglossal nucleus, the motor center controlling the tongue musculature, this projection could constitute a pathway for chemosensory information to influence tongue-flicking behavior. In summary, vomeronasal information is mostly relayed to the hypothalamus not via the nucleus sphericus but through other telencephalic structures. Convergence of olfactory and vomeronasal information appears to occur at different levels in the telencephalon. A neural substrate for the chemosensory control of tongue-flicking behavior is provided.

The vomeronasal system.

In addition to the main olfactory system, many vertebrates possess a vomeronasal system that conveys more specialized chemosensory information. Unlike the airborne, volatile stimuli detected by the main olfactory system, vomeronasal stimuli are typically proteins of the lipocalin family which bind small, volatile ligands. Despite the smaller number of vomeronasal receptor types, the projection patterns of the vomeronasal receptor neurons to multiple glomeruli in the accessory olfactory bulb appear to be more complicated than those of the main olfactory system. The vomeronasal system has a direct subneocortical projection to hypothalamic areas that mediates specific behavioural and hormonal responses to pheromonal stimuli. However, the integration and transmission of this information can be modulated by learning mechanisms. The aim of this article is to outline some of the functions of the vomeronasal system, and in particular to comment on recent advances in our understanding of how vomeronasal information is coded and processed.

Neural substrates for tongue-flicking behavior in snakes.

Snakes deliver odorants to the vomeronasal organ by means of tongue-flicks. The rate and pattern of tongue-flick behavior are altered depending on the chemical context. Accordingly, olfactory and vomeronasal information should reach motor centers that control the tongue musculature, namely, the hypoglossal nucleus (XIIN); however, virtually nothing is known about the circuits involved. In the present work, dextran amines were injected into the tongue of garter snakes (Thamnophis sirtalis) to identify the motoneurons of the XIIN. Tracers were then delivered into the XIIN to identify possible afferents of chemical information. Large injections into the XIIN yielded retrograde labeling in two chemosensory areas: the medial amygdala (MA) and the lateral posterior hypothalamic nucleus (LHN). Smaller injections only yielded labeled neurons in the LHN. In fact, the MA, which receives afferents from the accessory olfactory bulb, the rostroventral lateral cortex, and the nucleus sphericus, projects to the LHN. Injections into the MA did not show terminal labeling in the XIIN but in an area lateral to it. However, injections into the LHN gave rise not only to labeled fibers in the XIIN but also to retrograde labeling in the MA, thus confirming the chemosensory input to LHN. Injecting different fluorescent tracers into the tongue and into the LHN corroborated the projection from the LHN to the XIIN. The present report investigates further connections of the olfactory and vomeronasal systems and describes the afferent connections to XIIN in a nonmammalian vertebrate. The circuit for tongue-flicking behavior described herein should be evaluated using functional studies.

Afferent and efferent connections of the nucleus sphericus in the snake Thamnophis sirtalis: convergence of olfactory and vomeronasal information in the lateral cortex and the amygdala.

This paper is an account of the afferent and efferent projections of the nucleus sphericus (NS), which is the major secondary vomeronasal structure in the brain of the snake Thamnophis sirtalis. There are four major efferent pathways from the NS: 1) a bilateral projection that courses, surrounding the accessory olfactory tract, and innervates several amygdaloid nuclei (nucleus of the accessory olfactory tract, dorsolateral amygdala, external amygdala, and ventral anterior amygdala), the rostral parts of the dorsal and lateral cortices, and the accessory olfactory bulb; 2) a bilateral projection that courses through the medial forebrain bundle and innervates the olfactostriatum (rostral and ventral striatum); 3) a commissural projection that courses through the anterior commissure and innervates mainly the contralateral NS; and 4) a meager bilateral projection to the lateral hypothalamus. On the other hand, important afferent projections to the NS arise solely in the accessory olfactory bulb, the nucleus of the accessory olfactory tract, and the contralateral NS. This pattern of connections has three important implications: first, the lateral cortex probably integrates olfactory and vomeronasal information. Second, because the NS projection to the hypothalamus is meager and does not reach the ventromedial hypothalamic nucleus, vomeronasal information from the NS is not relayed directly to that nucleus, as previously reported. Finally, a structure located in the rostral and ventral telencephalon, the olfactostriatum, stands as the major tertiary vomeronasal center in the snake brain. These three conclusions change to an important extent our previous picture of how vomeronasal information is processed in the brain of reptiles.

1243 Electron microscopic study of the receptor cell regeneration in rat vomeronasal organ after nerve transection

In addition to the main olfactory system, many vertebrates have developed an accessory olfactory system (AOS). The AOS consists of the vomeronasal organ (VNO), a chemoreceptive structure situated at the base of the nasal septum which houses the vomeronasal sensory neurons; the accessory olfactory bulb (AOB), a structure of the forebrain that receives input from the VNO serving as the first processing center of vomeronasal information; and higher olfactory centers that receive direct or indirect information from the AOB. This chapter provides a general overview about the physiology and function of the VNO and AOB.

1244 Steroid 5alpha-reductase in the Wallerian degeneration of the rat sciatic nerve

In addition to the main olfactory system, many vertebrates have developed an accessory olfactory system (AOS). The AOS consists of the vomeronasal organ (VNO), a chemoreceptive structure situated at the base of the nasal septum which houses the vomeronasal sensory neurons; the accessory olfactory bulb (AOB), a structure of the forebrain that receives input from the VNO serving as the first processing center of vomeronasal information; and higher olfactory centers that receive direct or indirect information from the AOB. This chapter provides a general overview about the physiology and function of the VNO and AOB.

Zygomatico-maxillary fractures

In addition to the main olfactory system, many vertebrates have developed an accessory olfactory system (AOS). The AOS consists of the vomeronasal organ (VNO), a chemoreceptive structure situated at the base of the nasal septum which houses the vomeronasal sensory neurons; the accessory olfactory bulb (AOB), a structure of the forebrain that receives input from the VNO serving as the first processing center of vomeronasal information; and higher olfactory centers that receive direct or indirect information from the AOB. This chapter provides a general overview about the physiology and function of the VNO and AOB.