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: 9370779
• 负责人: Elizabeth Zuniga-Sanchez
• 金额: $ 10.75万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9370779
• 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; Neuraxis; Neurons; Pathway interactions; Phase; Phenotype; Photoreceptors; Positioning Attribute; Preparation; Process; Proteins; Public Health; Regulation; Research; Research Personnel; Resources; Rest; Retina; Retinal; Retinal Cone; Retinal Diseases; Role; Schedule; Signal Transduction; Specific qualifier value; Specificity; Synapses; System; Technology; 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; 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发育的中心问题。神经元如何发现 他们的正确目标仍然很大程度上未知。视网膜中的第一个突触提供了一个简单且 实验上易于处理的系统探索突触特异性的基础。杆状和锥状光感受器 分别选择性地使用杆双极和锥双极突触。确定分子机制 介导特异性，我生成了不同光感受器及其相应的转录组图谱 突触发生过程中双极细胞的靶点。从这些数据集中，我发现编码细胞表面的不同转录本 并分泌蛋白质作为该过程的有希望的候选者。利用体内基因操作和 发展方法中，我将评估这些基因在建立适当连接方面的功能。从 在我的初始屏幕中，我发现以下内容：1）杆和杆双极之间的非规范 Wnt 信号传导是 负责决定突触层的位置，2) 杆中的 Rspo2 参与层分离， 3) 视锥细胞中的 Nrcam 介导与视锥细胞双极的选择性连接。我把这个提案分为两部分 我在目标 1 中研究棒对棒双极连接，并在其中解决锥对锥双极连接 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相。第二部分涉及继续筛选和扩展新表型，这将是 在 S. Lawrence Zipursky 博士的指导下发起，并将在 R00 阶段完成。 该项目旨在帮助 Zuniga-Sanchez 博士获得终身教职助理职位 顶尖学术研究机构的教授。在导师和顾问委员会的帮助下，她 产生了有希望的数据并在她的研究中取得了重大进展。作为一名独立调查员， 她计划继续研究哺乳动物视网膜的发育，这与 人类健康。在K99阶段，她将继续受益于开发视觉效果的指导 Zipursky 博士的系统。她还将定期接受额外的职业和科学指导 与她的咨询委员会的会议和演讲。 Zuniga-Sanchez 博士将参加职业生涯 通过加州大学洛杉矶分校提供的发展研讨会，为申请教职和过渡到做准备 R00 阶段。该拨款提案概述了所需的培训、资源和合作网络 任命 Zuniga-Sanchez 博士为视觉领域的独立研究者。