Gbx2 regulates the development of an atypical amacrine cell.

Gbx2 调节非典型无长突细胞的发育。

• 批准号: 10221687
• 负责人: Patrick C Kerstein
• 金额: $ 1.34万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221687
• 关键词: Address; Adult; Amacrine Cells; Cells; Characteristics; Communication; Connexins; Coupled; Coupling; Data; Dendrites; Development; Disease; Electrical Synapse; Exhibits; Future; Gap Junctions; Gene Expression; Generations; Genes; Genetic; Genetic Identity; Genetic Transcription; Glycine; Health; Histology; Investigation; Knockout Mice; Knowledge; Label; Laboratories; Lead; Mediating; Molecular; Molecular Biology; Molecular Profiling; Morphogenesis; Morphology; Mus; Mutation; Neurons; Neurotransmitters; Output; Pathway interactions; Population; Process; Property; Regulation; Retina; Role; Stratification; Structure; Synapses; Testing; Tracer; Transgenic Mice; Vision; Visual system structure; base; cell type; conditional knockout; experimental study; gamma-Aminobutyric Acid; gene function; genetic approach; genetic manipulation; insight; motion sensitivity; neuron development; novel; protein expression; receptor; retinal neuron; selective expression; starburst amacrine cell; synaptogenesis; tool; transcription factor; transcriptome; transcriptome sequencing; visual information

PROJECT SUMMARY There are more than 100 distinct neuronal subtypes within the mammalian retina. Recent studies have made significant progress in our understanding of how specific neuron subtypes develop, wire into functional circuits, and contribute to disease. However, genetic tools to label and manipulate single populations of neurons only exist for a small proportion of retinal neurons. Using a genetic approach, I have identified a single population of retinal neurons based on their selective expression of the transcription factor Gbx2. These cells are amacrine cells (ACs) and have several interesting properties. Gbx2+ ACs do not express the inhibitory neurotransmitters, GABA or Glycine. Rather, Gbx2+ ACs exhibit gap junction coupled connections to bipolar cells, suggesting that the primary synaptic output for these ACs may be through electrical synapses. I will analyze the development of Gbx2+ ACs to investigate the role of Gbx2 in their cellular identity, morphology, and connectivity. In Aim 1, using a Gbx2 conditional knockout mouse, I will determine how Gbx2 regulates the morphology and electrical coupling of Gbx2+ ACs. I will also identify the specific connexins that mediate electrical synapse formation in Gbx2+ ACs. In Aim 2, I will perform RNAseq analyses in control and Gbx2 conditional knockout ACs to reveal their molecular profile and identify the specific effectors of Gbx2. I will also test the hypothesis that Gbx2 functions as a terminal fate selector to by regulating the expression of genes that endow Gbx2+ ACs with their unique characteristics. My preliminary data indicate that this occurs in part through the regulation of Robo receptors, which guide the proper stratification of Gbx2+ AC dendrites within the retina. This study will provided a comprehensive analysis of the morphological and molecular properties of a previously unidentified amacrine cell, and will elucidate how Gbx2 regulates the development and function of these neurons. The results of this study will lead to the generation of genetic tools and targets that will be useful for future investigations of retina development in health and disease.

项目摘要 哺乳动物视网膜内有100多个不同的神经元亚型。最近的 研究在我们对特定神经元亚型的理解方面取得了重大进步 发展，连接到功能电路，并导致疾病。但是，标记和标签的遗传工具 操纵神经元的单个群体仅在一小部分的视网膜神经元中存在。使用 遗传方法，我根据其选择性确定了单一的视网膜神经元种群 转录因子GBX2的表达。这些细胞是无结合细胞（AC），具有多个 有趣的属性。 GBX2+ ACS不会表达抑制性神经递质，GABA或甘氨酸。 相反，GBX2+ ACS表现出间隙连接与双极细胞的连接，表明该连接 这些AC的主要突触输出可能是通过电突触。我将分析 GBX2+ AC的开发，以研究GBX2在其细胞身份，形态和 连接性。在AIM 1中，使用GBX2条件敲除鼠标，我将确定GBX2的方式 调节GBX2+ ACS的形态和电耦合。我还将确定特定的 介导GBX2+ AC中介导电气突触形成的连接素。在AIM 2中，我将执行RNASEQ 对照和GBX2有条件敲除AC的分析，以揭示其分子轮廓并识别 GBX2的特定效应子。我还将测试GBX2作为终端命运的假设 通过调节将GBX2+ AC赋予其独特的基因表达来进行选择器 特征。我的初步数据表明，这部分是通过机器人的调节而发生的 受体，指导视网膜内GBX2+ AC树突的正确分层。这项研究会 提供了先前的形态和分子特性的全面分析 身份不明的裁活细胞，并将阐明GBX2如何调节 这些神经元。这项研究的结果将导致产生遗传工具和目标 对于对健康和疾病的视网膜发展的未来研究很有用。