Genetic Manipulation of Retinal Ganglion Cell Subtypes

项目摘要 光是一个非常重要的调节生理和行为在广泛的生物体。光 信息通过不同的视网膜神经节细胞类型传递到大脑中大约50个不同的目标。的 表达黑视素的内在光敏视网膜神经节细胞（ipRGC）代表40个视网膜神经节细胞中的6个。 存在于小鼠视网膜中的神经节细胞类型。M1-M6 ipRGC亚型各自具有不同的互补序列， 形态学和生理学特征、中枢投射和行为作用。因此，ipRGC的映射 具有单一子类型分辨率的电路将提供对多个和不同视觉功能的关键洞察， 电路.然而，这方面的进展一直受到缺乏标记，操纵， 和视网膜神经节细胞的单一亚型的消融。我们的初步数据表明，基于Cre和Flp的 交叉方法可以允许对单个ipRGC亚型进行这种操作。这项建议旨在 鉴定单个ipRGC亚型特有的遗传标记组合。同时，我们将 创建一个模块化的遗传工具包，可以整合到未来ipRGC亚型的任何Cre/Flp基序中 操纵这些工具不仅可以提供对ipRGC连接电路的即时洞察， 视觉和电路神经科学社区的关键工具和实验管道，以研究广泛的 将细胞功能与行为联系起来的一系列回路。

PROJECT SUMMARY Light is a profoundly important regulator of physiology and behavior across a wide range of organisms. Light information is relayed via diverse retinal ganglion cell types to approximately 50 distinct targets in the brain. The melanopsin-expressing, intrinsically photosensitive retinal ganglion cells (ipRGCs) represent 6 of the 40 retinal ganglion cell types present in the mouse retina. The M1-M6 ipRGC subtypes each possess a distinct complement of morphological and physiological features, central projections, and behavioral roles. Thus, mapping of ipRGC circuits with single subtype resolution would provide critical insight into the function of multiple and varied visual circuits. However, progress on this front has been hampered by a dearth of genetic tools for label, manipulation, and ablation of single subtypes of retinal ganglion cell. Our preliminary data indicate that a Cre and Flp-based intersectional approach could allow for such manipulation of single ipRGC subtypes. This proposal aims to identify genetic marker combinations unique to single ipRGC subtypes. Additionally, we will simultaneously create a modular genetic toolkit that can be integrated into any Cre/Flp motif for future ipRGC subtype manipulation. These tools will not only provide immediate insight into ipRGC-connected circuits, but will provide critical tools and an experimental pipeline for the visual and circuit neuroscience communities to study a broad range of circuits to link cellular function to behavior.