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.

Notch signaling determines cell-fate specification of the two main types of vomeronasal neurons of rodents.

The ability of terrestrial vertebrates to find food, mating partners and to avoid predators heavily relies on the detection of chemosensory information from the environment. The olfactory system of most vertebrate species comprises two distinct chemosensory systems usually referred to as the main and the accessory olfactory system. Olfactory sensory neurons of the main olfactory epithelium detect and transmit odor information to main olfactory bulb (MOB), while the chemosensory neurons of the vomeronasal organ detect semiochemicals responsible for social and sexual behaviors and transmit information to the accessory olfactory bulb (AOB). The vomeronasal sensory epithelium (VNE) of most mammalian species contains uniform vomeronasal (VN) system with vomeronasal sensory neurons (VSNs) expressing vomeronasal receptors of the V1R family. However, rodents and some marsupials have developed a more complex binary VN system, where VNO containing a second main type of VSNs expressing vomeronasal receptors of the V2R family is identified. In mice, V1R and V2R VSNs form from a common pool of progenitors but have distinct differentiation programs. As they mature, they segregate in different regions of the VNE and connect with different parts of the AOB. How these two main types of VSNs are formed has never been addressed. In this study, using single cell RNA sequencing data, we identified differential expression of Notch1 receptor and Dll4 ligand among the neuronal precursors at the VSN dichotomy. We further demonstrated with loss of function (LOF) and gain of function (GOF) studies that Dll4-Notch1 signaling plays a crucial role in triggering the binary dichotomy between the two main types of VSNs in mice.

Does a Third 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, Macropus 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, Macropus 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, which does not support the existence of a third “intermediate” type of vomeronasal information processing.

Effects of Amino Acids on Olfactory-Related Receptors Regulating Appetite in Silver Pomfret

Background: Amino acids are common components of the natural prey of fishes, and even single amino acid can bind olfactory receptors of fishes and induce appetite, which is regulated by hormones. Silver pomfret (Pampus argenteus) is one of the most preferred commercial marine fish species in many Asian countries and performs a significant appetite for jellyfish. Aspartic acid, glycine, and cysteine are high content amino acid in jellyfish, so we investigated the effect of three amino acids (aspartic acid, glycine, and cysteine) on olfactory-related receptors regulating appetite in silver pomfret (Pampus argenteus).Results: The fish showed significant attractant responses to these amino acids in water, which were also observed to improve the ingestion rates of the fish. Next, we conducted transcriptomes of the olfactory epithelium (OE) and identified 34 olfactory-related receptor genes were including olfactory receptors, trace amine-associated receptors and vomeronasal receptors genes, and we examined these genes in the OE and appetite-related genes (GHRL and LEP in gut/stomach and NPY, AgRP, and POMC in brain) by RT-qPCR. The olfactory-related receptor genes were significantly expressed in amino acid groups, and the appetite-related genes were most significantly expressed in aspartic acid group. Conclusions: Thus, olfactory-related receptors induced by amino acids might regulate appetite in silver pomfret through the OE-brain-gut/stomach axis. Using these data, we identified some effective amino acid phagostimulants which could be supplied in silver pomfret diet, and the results improved our understanding of the mechanism of olfactory-related receptors regulating appetite in fish.

Physiology-forward identification of bile acid–sensitive vomeronasal receptors

The mouse accessory olfactory system (AOS) supports social and reproductive behavior through the sensation of environmental chemosignals. A growing number of excreted steroids have been shown to be potent AOS cues, including bile acids (BAs) found in feces. As is still the case with most AOS ligands, the specific receptors used by vomeronasal sensory neurons (VSNs) to detect BAs remain unknown. To identify VSN BA receptors, we first performed a deep analysis of VSN BA tuning using volumetric GCaMP6f/s Ca2+ imaging. These experiments revealed multiple populations of BA-receptive VSNs with submicromolar sensitivities. We then developed a new physiology-forward approach for identifying AOS ligand-receptor interactions, which we call Fluorescence Live Imaging for Cell Capture and RNA sequencing, or FLICCR-seq. FLICCR-seq analysis revealed five specific V1R family receptors enriched in BA-sensitive VSNs. These studies introduce a powerful new approach for ligand-receptor matching and reveal biological mechanisms underlying mammalian BA chemosensation.

The Origin of Jaws and Paired Fins

Between 450 and 500 million years ago, some vertebrates evolved paired fins and jaws, which made them more efficient swimmers and fiercer predators. These jawed vertebrates (i.e., gnathostomes) diversified in the Devonian period, but most died out during the end-Devonian mass extinction. The surviving gnathostomes had a more complex vestibular apparatus than their jawless ancestors, an expanded set of olfactory receptor genes, and vomeronasal receptors. A major innovation in the brains of gnathostomes was the emergence of a cerebellum that is distinct from the cerebellum-like areas found in all vertebrates. The telencephalon of early vertebrates processed primarily olfactory information, but this olfactory dominance was independently reduced in three later lineages, namely in cartilaginous fishes, ray-finned fishes, and tetrapods. In concert with the reduction in olfactory dominance, these lineages enlarged their telencephalon, relative to other brain regions, and evolved a telencephalic “dorsal pallium” that receives non-olfactory sensory information from the diencephalon.

Physiology-forward identification of bile acid sensitive vomeronasal receptors

Abstract/SummaryThe mouse accessory olfactory system (AOS) supports social and reproductive behavior through the sensation of environmental chemosignals. A growing number of excreted steroids have been shown to be potent AOS cues, including bile acids (BAs) found in feces. As is still the case with most AOS ligands, the specific receptors used by vomeronasal sensory neurons (VSNs) to detect BAs remain unknown. To identify VSN BA receptors, we first performed a deep analysis of VSN BA tuning using volumetric GCaMP6f/s Ca2+ imaging. These experiments revealed both broadly and narrowly tuned populations of BA-receptive VSNs with sub-micromolar sensitivities. We then developed a new physiology-forward approach for identifying AOS ligand-receptor interactions, which we call Fluorescence Live Imaging for Cell Capture and RNA-seq, or FLICCR-seq. FLICCR-seq analysis revealed 5 specific V1R-family receptors enriched in BA-sensitive VSNs. These studies introduce a powerful new approach for ligand-receptor matching and reveal biological mechanisms underlying mammalian BA chemosensation.

HER2+ murine breast cancers and lung metastases differentially express a series of olfactory and vomeronasal receptors

I previously reported that over 15% of the most differentially expressed genes in CD24+ CD73+ γδ-T cells bearing a Vγ2 chain encode olfactory receptors. Here I present evidence that differential transcription of olfactory receptors can occur in breast cancers. Global comparative analysis of the transcriptomes of metastatic breast cancer in mice revealed that a series of Olfr (olfactory) genes were differentially expressed in tumors and in metastases. Lung metastases also expressed high levels of a non-coding RNA from the vomeronasal receptor family and two vomeronasal receptors. It is unknown whether any of these olfaction molecules play important roles in initiation or maintenance of tumors and/or metastases, or whether their expression can serve to improve diagnosis and/or prognosis. Differential expression of genes of the olfactory receptor family outside of the nervous system is not restricted to hematopoietic cells, and occurs in disease as well as in development.