The regulation of synaptic specificity in the mammalian retina

哺乳动物视网膜突触特异性的调节

• 批准号: 10179396
• 负责人: Elizabeth Zuniga-Sanchez
• 金额: $ 23.62万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10179396
• 关键词: Address; Advisory Committees; Applications Grants; Binding; Binding Proteins; Biological Assay; CRISPR/Cas technology; Cell Adhesion Molecules; Cell Surface Proteins; Cell surface; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Cone; Data; Data Set; Defect; Development; Educational workshop; Electroporation; Faculty; Gene Expression; Genes; Goals; Health; Human; Institution; Knock-out; Mediating; Mentors; Mentorship; Mission; Molecular; Mus; NRCAM gene; National Eye Institute; Neural Retina; Neuraxis; Neurons; Pathway interactions; Phase; Phenotype; Photoreceptors; Positioning Attribute; Preparation; Process; Proteins; Public Health; Regulation; Research; Research Personnel; Resources; Rest; Retina; Retinal Cone; Retinal Diseases; Rod; Role; Schedule; Signal Transduction; Specific qualifier value; Specificity; Synapses; System; Testing; Therapeutic; Tissue-Specific Gene Expression; Training; Transcript; Vertebrate Photoreceptors; Vision; Visual system structure; WNT Signaling Pathway; Work; base; career; career development; cell type; design; differential expression; experimental study; gain of function; gene function; genetic manipulation; in vivo; insight; meetings; neurodevelopment; novel; postsynaptic; presynaptic; professor; retinal rods; screening; segregation; sight restoration; synaptogenesis; tenure track; transcriptome; transcriptome sequencing; vision development

Abstract Establishing proper retinal circuitry remains a central issue in visual system development. How neurons find their correct target remains largely unknown. The first synapse in the retina provides a simple and experimentally tractable system to explore the basis of synaptic specificity. Rod and cone photoreceptors synapse selectively with rod bipolars and cone bipolars, respectively. To identify the molecular mechanism mediating specificity, I generated transcriptome profiles of the different photoreceptors and their corresponding bipolar cell targets during synaptogenesis. From these data sets, I find distinct transcripts encoding cell surface and secreted proteins as promising candidates for this process. Using in vivo genetic manipulations and developmental approaches, I will assess the function of these genes in establishing proper connectivity. From my initial screen, I find the following: 1) non-canonical Wnt signaling between rods and rod bipolars is responsible for dictating the position of the synaptic layer, 2) Rspo2 in rods is involved in layer segregation, and 3) Nrcam in cones mediates selective connectivity to cone bipolars. I subdivided this proposal into two parts where I study rod-to-rod bipolar connectivity in Aim 1 and address cone-to-cone bipolar connectivity in Aim 2. The first section of each aim involves expanding on my preliminary findings and will be completed in the K99 phase. The second part involves continue screening and expanding on the new phenotypes which will be initiated under the mentorship of Dr. S. Lawrence Zipursky and will be completed during the R00 phase. This project has been designed to facilitate Dr. Zuniga-Sanchez to obtain a position as a tenure-track Assistant Professor at a top-tier academic research institution. With the help of her mentor and Advisory Committee, she has generated promising data and made significant progress on her research. As an independent investigator, she plans to continue her work studying the development of the mammalian retina with a clear relevance to human health. During the K99 phase, she will continue benefiting from the mentorship on the developing visual system from Dr. Zipursky. She will also receive additional career and scientific guidance at regular scheduled meeting and presentations with her Advisory Committee. Dr. Zuniga-Sanchez will participate in career development workshops offered through UCLA in preparation to apply for faculty positions and transitioning to the R00 phase. This grant proposal outlines the training, resources, and network of collaborations needed to establish Dr. Zuniga-Sanchez as an independent investigator in the vision field.

抽象的 建立适当的视网膜电路仍然是视觉系统开发中的核心问题。神经元如何找到 他们的正确目标在很大程度上仍然未知。视网膜中的第一个突触提供了一个简单且 实验可拖动的系统，以探索突触特异性的基础。杆和锥形感受器 分别与杆双皮拉和锥双皮拉杆进行选择性突触。确定分子机制 介导的特异性，我生成了不同感光器及其相应的转录组曲线 突触发生过程中的双极细胞靶标。从这些数据集中，我发现编码细胞表面的不同成绩单 并分泌蛋白质作为此过程的有前途的候选人。使用体内遗传操作和 发展方法，我将评估这些基因在建立适当连通性方面的功能。从 我的初始屏幕，我找到以下内容：1）杆和杆双皮拉之间的非典型Wnt信号传导是 负责指示突触层的位置，2）杆中的rspo2参与层隔离， 3）锥体中的NRCAM介导了与锥双皮拉的选择性连通性。我将此提议分为两个 我在AIM 1中研究杆到杆双极连接的部分，并介绍锥形至圆锥形双极连通性 目标2。每个目标的第一部分涉及扩展我的初步发现，并将在 K99阶段。第二部分涉及继续筛选和扩展新表型 在S. Lawrence Zipursky博士的指导下启动，并将在R00阶段完成。 该项目旨在促进Zuniga-Sanchez博士获得任期助理的职位 顶级学术研究机构的教授。在她的导师和咨询委员会的帮助下，她 已经产生了有希望的数据，并在她的研究上取得了重大进展。作为独立研究者 她计划继续研究研究哺乳动物视网膜的发展，并与 人类健康。在K99阶段，她将继续从发展视觉的指导中受益 Zipursky博士的系统。她还将定期获得其他职业和科学指导 与她的咨询委员会会议和演讲。 Zuniga-Sanchez博士将参加职业 通过UCLA提供的开发研讨会准备申请教师职位并过渡到 R00阶段。该赠款提案概述了所需的培训，资源和合作网络 建立Zuniga-Sanchez博士为视觉领域的独立研究者。