Cytoskeletal regulation and Yap/Taz activity in differentiating retinal neurons

分化视网膜神经元中的细胞骨架调节和 Yap/Taz 活性

• 批准号: 9223832
• 负责人: Steven J Henle
• 金额: $ 10万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9223832
• 关键词: Address; Adhesions; Adult; Apical; Architecture; Back; Biological Assay; Biological Models; Blindness; Cell Differentiation process; Cell Nucleus; Cell division; Cells; Complement; Cytoskeleton; Defect; Development; Elements; Engineering; Feedback; Gatekeeping; Genetic Transcription; Goals; Human; Imaging Techniques; Injury; Knock-out; Knowledge; Lead; Link; Molecular; Molecular Genetics; Morphology; Mus; Mutation; Natural regeneration; Nerve Degeneration; Nerve Regeneration; Neuronal Differentiation; Neurons; Phase; Play; Process; Proteins; Regulation; Research; Retina; Retinal; Retinal Ganglion Cells; Role; Series; Side; Signal Pathway; Structural Protein; Surface; Systems Development; Technical Expertise; Techniques; Testing; Training; Transcriptional Activation; Transcriptional Regulation; Work; Zebrafish; cell type; experimental study; genetic manipulation; genetic regulatory protein; in vivo; in vivo imaging; innovation; insight; neurogenesis; neuron development; novel; prevent; progenitor; regenerative; regenerative therapy; repaired; retinal neuron; retinal progenitor cell; retinal regeneration; skills; tool; tool development

PROJECT SUMMARY / ABSTRACT The human retina does not regenerate after injury. This proposal examines retinal development to better understand the molecular mechanisms that could be used to initiate regeneration. Developing neurons must precisely regulate cytoskeletal dynamics to establish their unique morphology that is required for proper function. It is not well understood how cytoskeletal dynamics change across development and after injury in vivo. Yes Associated Protein (YAP) and Tafazzin (TAZ) are functionally similar proteins that are critical for maintaining the progenitor state through transcriptional regulation. Mutations to YAP/TAZ result in retinal lamination defects through an unknown mechanism. Additionally, unlike other retinal neurons, retinal ganglion cells continue to express YAP/TAZ after development. Intriguingly, cytoskeletal tension can inhibit YAP/TAZ activity and YAP/TAZ can regulate cytoskeletal dynamics. This creates an unusual molecular feedback loop between structural proteins and transcriptional regulators. The overall goal of this proposal is to understand the molecular mechanisms by which the cytoskeleton and YAP/TAZ coordinate retinal development, and the role they play in retinal regeneration. The central hypothesis is that cytoskeletal rearrangements required for retinal differentiation downregulate YAP/TAZ activity and thus drive cells out of the progenitor state. With the knowledge gained from these studies, the ultimate goal is to reverse engineer this process in adult cells, so we can then drive cells to a state of enhanced regenerative potential. To this end I will use a combination of innovative in vivo imaging techniques, genetic manipulations, and regeneration assays to better understand the molecular mechanisms by which the cytoskeleton and YAP/TAZ orchestrate retinal development and regeneration. The initial experiments in this proposal will focus on developing tools and techniques in zebrafish to investigate the cytoskeleton in order to test the overall hypothesis. Mastery of the zebrafish model system is a key element of the training potential of the K99 phase, and will complement my previous work examining retinal development in mice. Having technical expertise in both mouse and zebrafish will enable me to answer the most critical questions by having experimental access across retinal development, and a wide range of molecular and genetic tools. Taken together this training plan will empower me to develop an independent line of research that can significantly contribute to the development of regenerative therapies for the retina, and ultimately prevent vision loss.

项目摘要/摘要 人的视网膜在受伤后不会再生。这项建议检查视网膜发育以更好地 了解可用于启动再生的分子机制。发育中的神经元必须 精确地调节细胞骨架动力学，以建立其独特的形态，这是正常 功能。目前还不清楚细胞骨架动力学如何在发育过程中和受伤后发生变化。 活着。是的，相关蛋白(YAP)和他法津(TAZ)是功能相似的蛋白质，对 通过转录调控维持祖细胞状态。YAP/TAZ突变导致视网膜 层压缺陷通过一种未知的机制。此外，与其他视网膜神经元不同，视网膜神经节 细胞发育后继续表达YAP/TAZ。有趣的是，细胞骨架张力可以抑制YAP/TAZ 活性和YAP/TAZ对细胞骨架动力学有调节作用。这就产生了一个不同寻常的分子反馈回路 结构蛋白和转录调控因子之间的关系。这项建议的总体目标是理解 细胞骨架和YAP/TAZ协调视网膜发育的分子机制，以及 它们在视网膜再生中扮演着重要角色。中心假设是细胞骨架重排所需的 视网膜分化下调YAP/TAZ的活性，从而将细胞赶出祖细胞状态。与 从这些研究中获得的知识，最终目标是在成年细胞中对这一过程进行反向工程，所以我们 然后可以将细胞驱动到增强的再生潜力的状态。为此，我将结合使用 创新的活体成像技术、基因操作和再生分析，以更好地了解 细胞骨架和YAP/TAZ调控视网膜发育的分子机制 再生。 这项计划中的初步实验将集中于开发斑马鱼的工具和技术，以研究 细胞骨架，以检验整体假说。掌握斑马鱼模型系统是一个关键因素 K99阶段的训练潜力，并将补充我之前检查视网膜的工作 在小鼠体内发育。在老鼠和斑马鱼方面都有技术专长将使我能够回答 最关键的问题，通过在视网膜发育过程中进行试验性访问，以及广泛的 分子和遗传工具。综上所述，这个培训计划将使我有能力发展一条独立的路线 可以对视网膜再生疗法的开发做出重大贡献的研究，以及 最终防止视力丧失。