Glomerular specific neuromodulation via differential serotonin receptor trafficking

通过差异血清素受体运输进行肾小球特异性神经调节

• 批准号: 10043109
• 负责人: Quentin Gaudry
• 金额: $ 20.83万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-12 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10043109
• 关键词: Address; Adopted; Anatomy; Antibodies; Area; Biological Models; Brain; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Communities; Dendrites; Drosophila genus; Electrophysiology (science); Funding; Genetic; Genetic Recombination; Genome; Green Fluorescent Proteins; Immunohistochemistry; Individual; Interneurons; Label; Libraries; Lobe; Mediating; Microscopy; Mission; Modeling; Monitor; Mus; National Institute on Deafness and Other Communication Disorders; Neuromodulator; Neurons; Neuropil; Nociception; Odors; Olfactory Pathways; Output; Pattern; Population; Process; Reagent; Site; Smell Perception; Surveys; Synapses; System; Taste Perception; Testing; Visualization; Work; analog; cell type; experimental study; fly; mitral cell; nerve supply; neuroregulation; novel; olfactory bulb; olfactory bulb glomeruli; postsynaptic; presynaptic; receptor; response; sensory system; serotonin receptor; tool; trafficking; virtual

This proposal examines the contribution of receptor trafficking in inhibitory interneurons for establishing glomerulus-specific neuromodulation during olfactory. Current models of neuromodulation in olfaction emphasize the activation of specific classes of neurons that express a specific and dedicated class of neuromodulatory receptor, for example a single serotonin receptor. In such models, local interneurons in the first stages of olfactory processing may differentially innervate distinct olfactory glomeruli depending on the modulatory receptors that they express. Here, we will challenge this view and develop tools to demonstrate that odor specific modulation may occur not solely by targeting distinct cell classes, but also by the dendrite specific modulation of local interneurons that span multiple olfactory glomeruli. Specifically, we will develop tools to tag and track each serotonin receptor type in identified classes of local interneurons in the Drosophila antennal lobe, the first olfactory relay. Additionally, we will tag and label serotonin receptors in projection neurons that span multiple olfactory neuropil to determine how serotonin receptors are trafficked across olfactory neuropil. The Drosophila antennal lobe is an excellent model system for addressing these issues as its organization is highly similar to the mammalian olfactory bulb, and the smaller number of cells in the antennal lobe render it easier to study and characterize. Receptor tagging will be achieved be editing the fly genome to express modified serotonin receptors that are fused with the 11th fragment of the green fluorescent protein molecule, GFP. A genetic binary expression system can then be used to express the remaining portion of the split-GFP molecule to selectively label serotonin receptors in a specific cell class or individual neuron. We combine these tools with expansion microcopy to examine the co-localization of serotonin receptors with presynaptic and postsynaptic release sites within local interneuron and projection neurons populations. Finally, we will generate a version of these tools using a split mCherry fluorescent molecule approach so that the co-localization of serotonin receptor classes can be examined when local interneurons express multiple classes of these receptors. These experiments will lay the anatomical ground work for subsequent studies examining functional branch-specific neuromodulation of wide-field local interneurons in early olfactory processing.

这项建议研究了抑制性中间神经元中受体运输的贡献 在嗅觉过程中建立肾小球特异性神经调节。当前型号的 嗅觉中的神经调节强调激活特定类别的神经元，这些神经元 表达一类特定的和专用的神经调节受体，例如单个 5-羟色胺受体在这样的模型中，嗅觉处理第一阶段的局部中间神经元 可能根据调制受体不同而不同地支配不同的嗅觉小球 他们所表达的。在这里，我们将挑战这一观点，并开发工具来演示这种气味 特定的调制不仅可以通过针对不同的细胞类别来进行，还可以通过 跨越多个嗅觉小球的局部中间神经元的树突特异性调节。 具体地说，我们将开发工具来标记和跟踪已识别的每一种血清素受体类型 果蝇触角叶局部中间神经元的分类，第一嗅觉中继器。 此外，我们将标记和标记横跨多个 以确定5-羟色胺受体是如何通过嗅神经管运输的。 果蝇的触角叶是解决这些问题的一个很好的模型系统，因为它的 组织结构与哺乳动物的嗅球高度相似，而且细胞数量较少 触角叶使其更容易研究和表征。将实现受体标记 正在编辑苍蝇基因组以表达与第11条融合的修饰的5-羟色胺受体 绿色荧光蛋白分子GFP的片段。一种遗传二进制表达系统 然后可以用来表达分裂的GFP分子的剩余部分来选择性地标记 特定细胞类别或单个神经元中的5-羟色胺受体。我们将这些工具与 扩张显微镜检查5-羟色胺受体与突触前和突触前的共定位 局部中间神经元和投射神经元群体中的突触后释放部位。最后， 我们将使用分裂的mCherry荧光分子方法生成这些工具的一个版本 这样就可以在局部检查5-羟色胺受体类别的共同定位 中间神经元表达多种类型的这些受体。这些实验将为 检查特定功能分支的后续研究的解剖学基础工作 早期嗅觉加工中广域局部中间神经元的神经调节。