An ex vivo preparation of the intact mouse vomeronasal organ and accessory olfactory bulb

Meeks, Julian P.; Holy, Timothy E.

doi:10.1016/j.jneumeth.2008.11.013

Cited by 15 publications

(34 citation statements)

References 32 publications

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“…However, studies in zebrafish and mice have previously utilized similar ex vivo preparations in Ringer's solution in order to examine olfactory responses in the olfactory epithelium, main and accessory olfactory bulbs, and higher brain structures, with spatial organization of odour information similar to that reported in vivo Korsching, 1997, 1998;Meeks and Holy, 2009). In this ex vivo preparation, the lateral region of the lamprey olfactory bulb was a locus for amino acid responses and the dorsal region of the sea lamprey olfactory bulb responded to the steroid bile acid taurocholic acid in all cases, to both taurocholic acid and the steroid pheromones in 30% of cases, and to amino acids, taurocholic acid and pheromones in 50% of cases, and was the only region with cases that responded to taurocholic only (20%).…”

Section: Discussionmentioning

confidence: 99%

Odorant organization in the olfactory bulb of the sea lamprey

Green

Boyes

McFadden

et al. 2017

Journal of Experimental Biology

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Olfactory sensory neurons innervate the olfactory bulb, where responses to different odorants generate a chemotopic map of increased neural activity within different bulbar regions. In this study, insight into the basal pattern of neural organization of the vertebrate olfactory bulb was gained by investigating the lamprey. Retrograde labelling established that lateral and dorsal bulbar territories receive the axons of sensory neurons broadly distributed in the main olfactory epithelium and that the medial region receives sensory neuron input only from neurons projecting from the accessory olfactory organ. The response duration for local field potential recordings was similar in the lateral and dorsal regions, and both were longer than medial responses. All three regions responded to amino acid odorants. The dorsal and medial regions, but not the lateral region, responded to steroids. These findings show evidence for olfactory streams in the sea lamprey olfactory bulb: the lateral region responds to amino acids from sensory input in the main olfactory epithelium, the dorsal region responds to steroids (taurocholic acid and pheromones) and to amino acids from sensory input in the main olfactory epithelium, and the medial bulbar region responds to amino acids and steroids stimulating the accessory olfactory organ. These findings indicate that olfactory subsystems are present at the base of vertebrate evolution and that regionality in the lamprey olfactory bulb has some aspects previously seen in other vertebrate species.

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Section: Discussionmentioning

confidence: 99%

Odorant organization in the olfactory bulb of the sea lamprey

Green

Boyes

McFadden

et al. 2017

Journal of Experimental Biology

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“…There is evidence for gradual degeneration of the deepest AOB layers, namely the deep portions of the internal cellular layer, which houses many inhibitory granule cells 16 . Additionally, centrifugal inputs are severed during the dissection, eliminating them from active participation in AOB function.…”

Section: Discussionmentioning

confidence: 99%

“…Because the connections between the VNO and AOB remain ipsilateral, and because the midline septal tissue can be exposed to oxygenated superfusate in a single hemisphere, we sought to develop such a single-hemisphere ex vivo approach to isolate these structures while maintaining their functional connectivity. We recently succeeded in achieving this goal 16 . This preparation keeps both the VNO and AOB alive and functionally connected for at least 4 -6 hr because both the axons (along the midline soft septal tissue) and AOB are relatively shallow <600 µm features that are accessible to superfused oxygenated artificial cerebrospinal fluid (aCSF).…”

Section: Introductionmentioning

confidence: 99%

<em>Ex Vivo</em> Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Doyle

Hammen

Meeks

2014

JoVE

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The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first neural circuit in the AOS pathway, called the accessory olfactory bulb (AOB), plays an important role in establishing sextypical behaviors such as territorial aggression and mating. This small (<1 mm 3 ) circuit possesses the capacity to distinguish unique behavioral states, such as sex, strain, and stress from chemosensory cues in the secretions and excretions of conspecifics. While the compact organization of this system presents unique opportunities for recording from large portions of the circuit simultaneously, investigation of sensory processing in the AOB remains challenging, largely due to its experimentally disadvantageous location in the brain. Here, we demonstrate a multi-stage dissection that removes the intact AOB inside a single hemisphere of the anterior mouse skull, leaving connections to both the peripheral vomeronasal sensory neurons (VSNs) and local neuronal circuitry intact. The procedure exposes the AOB surface to direct visual inspection, facilitating electrophysiological and optical recordings from AOB circuit elements in the absence of anesthetics. Upon inserting a thin cannula into the vomeronasal organ (VNO), which houses the VSNs, one can directly expose the periphery to social odors and pheromones while recording downstream activity in the AOB. This procedure enables controlled inquiries into AOS information processing, which can shed light on mechanisms linking pheromone exposure to changes in behavior. Video LinkThe video component of this article can be found at

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“…Ex vivo Ca 2+ imaging and data analysis. Glomerular imaging was performed using a VNO-AOB ex vivo preparation (Hammen et al, 2014;Meeks and Holy, 2009). Briefly, after deep isofluorane anesthesia, mice were decapitated and placed into ice-cold aCSF.…”

Section: Methodsmentioning

confidence: 99%

Physiology-forward identification of bile acid sensitive vomeronasal receptors

Wong

Cao

Zhang

et al. 2019

Preprint

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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 Ca 2+ 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. Introduction:Chemosensory systems extract salient information from environmental cues that are critical for social and reproductive behaviors. In mice and other terrestrial mammals, the accessory olfactory system (AOS) guides innate behaviors such as territorial aggression and mating (Dulac and Torello, 2003;Keller et al., 2009). The initial detection of olfactory stimuli in the AOS is mediated by vomeronasal sensory neurons (VSNs), located in the vomeronasal organ (VNO), before these signals are routed to the accessory olfactory bulb and then behaviorally important regions including the medial amygdala and bed nucleus of the stria terminalis.Though the behavioral impacts of AOS chemosensation are manifold, our knowledge of the full complement of natural ligands for VSNs remains incomplete. Natural AOS ligands are found in the excretions of conspecific and heterospecific animals (e.g., urine, feces, tears, and saliva). These natural ligands include, but are not limited to, polar steroids, bile acids, major urinary proteins, and major histocompatibility complex peptide ligands (Chamero et al., 2007;Doyle et al., 2016;Leinders-Zufall et al., 2004;Nodari et al., 2008). As we learn more about the full natural complement of AOS ligands, new questions are arising about how these cues are detected by specific types of VSNs. Such knowledge is critical as we seek to understand how patterns of VSN activation by complex blends of environmental chemosignals cause changes in animal physiology and behavior.A major barrier preventing a deeper understanding of AOS function is a lack of ligandreceptor matches for AOS cues. VSNs generally express just one or two of ~300 unique vomeronasal receptors (VRs) or formyl peptide receptors (Dulac and Torello, 2003;Martini et al., 2001). The largely monoallelic expression of individual VRs by VSNs means that each VSN takes on the chemosensory sensitivity (or "tuning") of the dominantly expressed receptor. In theory, this feature wou...

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An ex vivo preparation of the intact mouse vomeronasal organ and accessory olfactory bulb

Cited by 15 publications

References 32 publications

Odorant organization in the olfactory bulb of the sea lamprey

Odorant organization in the olfactory bulb of the sea lamprey

<em>Ex Vivo</em> Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Physiology-forward identification of bile acid sensitive vomeronasal receptors

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