Molecular and cellular analysis of accessory olfactory circuits in mice

小鼠辅助嗅觉回路的分子和细胞分析

• 批准号: 9816360
• 负责人: Gilad Barnea
• 金额: $ 4.17万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9816360
• 关键词: Address; Affect; Afferent Neurons; Animals; Antibodies; Apical; Axon; Behavior; Behavioral; Biochemical; CRISPR/Cas technology; Caring; Chemicals; Chromosomes; Chromosomes, Human, Pair 3; Complex; Continuing Education; Detection; Distant; Engineering; Environment; Exhibits; Extracellular Domain; Family; Foundations; Future; G-Protein-Coupled Receptors; Gene Cluster; Genes; Genetic Recombination; Glutamates; Human; Immunoprecipitation; Insecta; Instinct; Knock-out; Ligand Binding; Ligands; Light; Location; Lordosis; Mass Spectrum Analysis; Mediating; Molecular; Mus; Neurons; Odorant Receptors; Odors; Olfactory Epithelium; Olfactory Pathways; Organ; Partner in relationship; Pattern; Personal Satisfaction; Pheromone; Property; Proteins; Reagent; Research; Research Personnel; Rest; Role; Schools; Sensory; Signal Pathway; Signal Transduction; Social Behavior; Social Controls; Solid; Stratum Basale; Supervision; Taste Buds; Techniques; Territoriality; Testing; Training; Underrepresented Minority; Western Blotting; Yang; behavioral response; experience; experimental study; genetic makeup; human disease; insight; member; mouse genome; multidisciplinary; neural circuit; neuromechanism; olfactory receptor; receptor; relating to nervous system; response; sweet receptor; training opportunity; transcriptome sequencing; vomeronasal organ

In mice, the vomeronasal organ (VNO) is a main sensory organ for detecting pheromones, chemicals that affect social behaviors including: territoriality, sexual recognition and maternal care. The VNO can be divided into apical and basal layers that differ in their properties. VNO sensory neurons (VSNs) in the apical layer express members of the Vmn1r family of G protein coupled receptors (GPCRs), while basal VSNs express members of the Vmn2r family of GPCRs. Vmn2rs belong to the same family of GPCRs that includes the metabotropic glutamate, GABAB, and taste receptors. Basal VSNs do not obey the one receptor per neuron rule that is operative in most other sensory neurons in the olfactory system. The Vmn2r family consists of four classes designated A, B, C and D. Members of classes A, B and D are more closely related to one another than to the seven members of class C (Vmn2r1-7). Each basal VSN expresses one Vmn2rC and one Vmn2rABD. Consequently, The seven class C receptors are broadly expressed in the VNO while class ABD receptors are sparsely expressed, like the rest of the olfactory receptors. The functional significance of the coexpression of Vmn2rs is not well understood. We hypothesize that Vmn2rCs and Vmn2rABDs form heterodimers and that this interaction alters the functional properties of the basal VSNs. Vmn2rCs might affect the subcellular localization of Vmn2rABDs, their ligand binding and signaling properties. To examine these hypotheses, we have devised a multipronged strategy encompassing molecular and biochemical studies, neuroanatomical examination, and behavioral analysis. We have generated a mouse line carrying a deletion of the Vmn2r1-7 gene cluster using CRISPR/Cas9-mediated chromosome engineering followed by Cre recombination. Further, we generated a battery of specific antibodies against all Vmn2rCs, against specific class C receptors, and against a clade of Vmn2rAs. With these new reagents, we are poised to address these hypotheses. Our studies will reveal the mechanisms underlying the function of basal VSNs. More broadly, these experiments will shed light on how the olfactory system mediates social behaviors that are critical for the survival of the species. Thus far, only a handful of Vmn2r ligands have been identified. Since we predict that the deletion of the Vmn2rC cluster will affect the ability of the animals to properly respond to Vmn2rABD ligands, the cluster knockout line that we have generated will be invaluable for evaluating new Vmn2r ligands and their behavioral importance, as these ligands are identified. In this manner, our studies will deepen our understanding of the control of social behavior in multiple ways. Finally, our experiments will provide additional insight into the molecular mechanisms underlying the function of the GPCR family to which Vmn2rs belong. Some of these receptors have relevance to human disease. This project will offer Tariq Cannonier, a recent Brown graduate from an under-represented minority, a unique training opportunity that will provide him a solid foundation for applying to graduate school for his continuing education.

在小鼠中，呕吐器官（VNO）是检测信息素的主要感觉器官，化学物质 影响社会行为，包括：领土，性识别和孕产妇护理。 VNO可以分开 变成其性质不同的顶端和基础层。顶层的VNO感觉神经元（VSN） G蛋白偶联受体（GPCR）的VMN1R家族的表达成员，而基底VSN表示 GPCR的VMN2R家族的成员。 VMN2RS属于包括包括的GPCR家族 代谢性谷氨酸，GABAB和味觉受体。基底VSN不遵守每个神经元的一个受体 在嗅觉系统中大多数其他感觉神经元中起作用的规则。 VMN2R家族由四个 指定A，B，C和D的类 比C类（VMN2R1-7）的七个成员。每个基底VSN表示一个VMN2RC和一个 VMN2RABD。因此，七个C类受体在VNO中广泛表达 像其他嗅觉受体一样，受体表达稀少。功能意义的 VMN2R的共表达尚不清楚。我们假设VMN2RC和VMN2RABD形式 异二聚体，这种相互作用会改变基底VSN的功能特性。 VMN2RC可能会影响 VMN2RABD的亚细胞定位，其配体结合和信号传导特性。 为了审查这些假设，我们制定了一种涉及分子和 生化研究，神经解剖学检查和行为分析。我们已经生成了一条鼠标 使用CRISPR/CAS9介导的染色体工程删除VMN2R1-7基因群 其次是CRE重组。此外，我们生成了针对所有VMN2RC的一系列特定抗体， 针对特定的C类受体，并针对VMN2RA的进化枝。有了这些新试剂，我们准备 解决这些假设。我们的研究将揭示基础VSN功能的基础机制。 从更广泛的角度来看，这些实验将阐明嗅觉系统如何介导的社会行为 对物种的生存至关重要。到目前为止，仅确定了少数VMN2R配体。自从我们 预测VMN2RC群集的删除将影响动物正确响应的能力 VMN2RABD配体，我们生成的集群敲除线对于评估新的是无价的 VMN2R配体及其行为重要性，因为这些配体被确定。这样，我们的研究将 加深我们以多种方式对社会行为控制的理解。最后，我们的实验将 对GPCR家族功能的基础分子机制提供更多的见解 vmn2rs属于。这些受体中的一些与人类疾病有关。这个项目将提供Tariq 坎诺尼尔（Cannonier），最近来自代表性不足的少数民族的棕色毕业生，这是一个独特的培训机会 为他为继续教育的研究生院申请稳固的基础。