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.

项目概要 视网膜是研究神经回路发育和功能的绝佳模型。这 视网膜包含五个主要的神经元类别，可细分为一百多个神经元 亚型。无长突细胞 (AC) 是五种神经元类型之一，仅占视网膜细胞的百分之一 人口，但约占该亚型多样性的一半。仅少数 AC 亚型已被证实 进行了深入研究，部分原因是缺乏前瞻性隔离和操纵个体群体的工具。 因此，我们对视觉回路如何运作的理解仍然不完整。比如最近的工作 表明方向选择电路是视网膜中研究最深入的电路之一，包括甘氨酸 来自未定义的 AC 子类型的输入。要充分了解 AC 如何促进视觉处理，需要 迫切需要研究更多的 AC 亚型及其定义特征。 在初步研究中，我发现了一个 Robo3CreER 小鼠品系，该品系对一群 以前未描述过的甘氨酸无长突细胞。根据它们的形态和分层模式，我 假设这些 Robo3+ 无长突细胞 (RAC) 可能是对 ON 提供甘氨酸抑制的细胞 方向选择电路中的星爆无长突细胞。我将采用小鼠遗传学、共焦显微镜 原位杂交、电子显微镜和电生理学来检验这一假设。在目标 1 中，我将定义 RAC 的形态特性和群体水平组织。在目标 2 中，我将确定 RAC 是否 在星爆无长突细胞上形成抑制性突触。在目标 3 中，我将确定 Robo3 是否扮演 在 RAC 开发和/或功能中的作用。这些目标将拓宽我们对个体细胞如何 视网膜电路中的各个组件一起发挥作用。