Novel Activators of Regeneration in Muller glia

项目概要：新型Müller胶质细胞再生激活剂 Müller神经胶质细胞是鱼类视网膜再生修复的主要来源，但在视网膜中并非如此。 在人类中，Müller神经胶质通过变得反应性而对损伤、病理或变性作出反应，并且在许多情况下， 例胶质瘤。最近对非再生哺乳动物模型系统（如小鼠）的研究表明， 米勒神经胶质细胞可以被推下再生途径，但真正的再生和修复将 涉及尚未确定的因素和协议。为了弥补这一知识差距，我们提出三点建议。 这些项目侧重于再生所需的增殖和神经原性。在第一个项目中， 我们将通过调节神经递质来开发小鼠Müller胶质细胞的再生启动方案 p27 Kip 1是一种细胞周期蛋白，可阻止与反应性相关的特性， Müller胶质细胞再生。在第二个项目中，我们将使用外泌体进行筛选， 候选外泌体和它们的组成蛋白和/或RNA，其刺激细胞的再生特性， 斑马鱼和小鼠Müller胶质细胞。在第三个项目中，我们将开发与RNA结合的硫氧嘧啶标签 测序以捕获基于我们的工作的Müller胶质细胞在选定再生状态下的转录组 前两个项目。这项工作的成果将产生促进再生的新因素， 为基于外泌体的因子递送作为治疗策略提供概念验证。

Project Summary: Novel Activators of Regeneration in Müller Glia Müller glia are the predominant source of regenerative repair in the fish retina, but this is not the case in humans, where Müller glia respond to injury, pathology, or degeneration by becoming reactive, and, in many cases, gliotic. Recent research in non-regenerative mammalian model systems, such as the mouse, suggests that Müller glia could be pushed down a regenerative pathway, but bona-fide regeneration and repair will involve factors and protocols that are not yet identified. To address this gap in knowledge, we propose three projects that focus on the proliferative and neurogenic properties needed for regeneration. In the first project, we will develop a regeneration-priming protocol for mouse Müller glia through modulation of neurotransmitter uptake and inhibition of p27Kip1, a cell cycle protein that prevents properties associated with reactivity and regeneration in Müller glia. In the second project, we will perform a screen using exosomes to discover candidate exosomes and their constituent proteins and/or RNAs that stimulate regenerative properties in zebrafish and mouse Müller glia. In the third project, we will develop thiouracil tagging combined with RNA sequencing to capture the transcriptomes of Müller glia in selected states of regeneration based on our work from the first two projects. The culmination of this work will yield new factors for promoting regeneration and provide proof-of-concept for exosome-based delivery of factors as a therapeutic strategy.