Genetic analysis of the Robo3+ glycinergic amacrine cell

Robo3 甘氨酸无长突细胞的遗传分析

• 批准号: 10749795
• 负责人: Arielle Isakharov
• 金额: $ 5.02万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2026-09-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749795
• 关键词: Affect; Amacrine Cells; Antibodies; Biological Models; Biotin; Candidate Disease Gene; Cell Separation; Cell Survival; Cells; Characteristics; Confocal Microscopy; Data; Data Set; Dendrites; Development; Electron Microscopy; Electrophysiology (science); Genetic; Gold; Growth; In Situ Hybridization; Individual; Inhibitory Synapse; Inner Plexiform Layer; Knock-out; Knowledge; Label; Membrane; Mitosis; Molecular Profiling; Morphology; Mus; Neurons; Neurophysiology - biologic function; Pattern; Population; Population Distributions; Population Sizes; Property; Reporter; Resolution; Retina; Shapes; Specific qualifier value; Stratification; Structure; Study models; Synapses; Testing; Time; Visual; Work; axon guidance; density; experimental study; falls; genetic analysis; genetic approach; inhibitory neuron; mouse genetics; multimodality; nervous system development; neural circuit; patch clamp; postnatal; prospective; receptor; response; retinal neuron; selective expression; single-cell RNA sequencing; starburst amacrine cell; synaptogenesis; tool; visual processing

Project Summary The retina is an excellent model for studying the development and function of neural circuitry. The retina contains five major neuronal classes which can be subdivided into well over a hundred neuronal subtypes. Amacrine Cells (ACs), one of the five neuronal classes, comprise only one percent of the retinal cell population, but account for around half of this subtype diversity. Only a handful of AC subtypes have been studied in depth, in part due to a lack of tools to prospectively isolate and manipulate individual populations. Therefore, our understanding of how visual circuits function remains incomplete. For example, recent work shows that the direction selective circuit, one of the most well-studied circuits in the retina, includes glycinergic input from an undefined AC subtype. To fully understand how ACs contribute to visual processing it is imperative to study more AC subtypes and their defining features. In preliminary studies, I identified a Robo3CreER mouse line that genetically labels a population of previously undescribed glycinergic amacrine cells. Based on their morphology and stratification pattern, I hypothesize that these Robo3+ amacrine cells (RACs) may be the cells that supply glycinergic inhibition to ON starburst amacrine cells in the direction selective circuit. I will employ mouse genetics, confocal microscopy, in situ hybridization, electron microscopy, and electrophysiology to test this hypothesis. In Aim 1, I will define the morphological properties and population-level organization of RACs. In Aim 2, I will determine whether RACs form inhibitory synapses onto ON starburst amacrine cells. In Aim 3, I will determine whether Robo3 plays a role in RAC development and/or function. These aims will broaden our knowledge of how individual cellular components function together in retinal circuitry.

项目摘要 视网膜是研究神经回路的发展和功能的绝佳模型。这 视网膜包含五个主要的神经元类别，可以细分为一百多个神经元 亚型。五个神经元类别之一的小氨基细胞（ACS）仅占视网膜细胞的百分之一 人口，但约占这种亚型多样性的一半。只有少数AC亚型 深入研究，部分原因是缺乏前瞻性隔离和操纵个体人群的工具。 因此，我们对视觉电路如何功能的理解仍然不完整。例如，最近的工作 表明选择性电路是视网膜中最有研究的电路之一，包括甘油能 输入未定义的AC亚型。为了充分了解ACS如何为视觉处理做出贡献 必须研究更多的AC亚型及其定义特征。 在初步研究中，我确定了一条Robo3creer小鼠系，该系列遗传标记了一个人群 以前未描述的甘霉素无长链氨酸细胞。根据它们的形态和分层模式，我 假设这些Robo3+无氨基氨酸细胞（RAC）可能是提供甘氨酸能抑制的细胞 Starburst amacrine细胞沿方向选择性电路。我将在中使用小鼠遗传学，共聚焦显微镜 原位杂交，电子显微镜和电生理学，以检验该假设。在AIM 1中，我将定义 RAC的形态学特性和人口水平的组织。在AIM 2中，我将确定RAC是否 形成抑制性突触在星爆肿瘤细胞上。在AIM 3中，我将确定Robo3是否扮演 在RAC开发和/或功能中的作用。这些目标将扩大我们对个体细胞如何的了解 组件在视网膜电路中共同发挥作用。