Genetic Manipulation of Retinal Ganglion Cell Subtypes

项目总结 光对各种生物体的生理和行为都起着极其重要的调节作用。灯 信息通过不同类型的视网膜神经节细胞传递到大脑中大约50个不同的目标。这个 表达黑色素的固有的光敏性视网膜神经节细胞(IpRGCs)代表40个视网膜中的6个 小鼠视网膜中存在神经节细胞类型。M1-M6 ipRGC亚型各自具有不同的补体 形态和生理特征、中央投射和行为作用。因此，ipRGC的映射 具有单个子类型分辨率的电路将提供对多种不同视觉功能的重要洞察 电路。然而，由于缺乏用于标记、操纵 和单一亚型视网膜神经节细胞的消融。我们的初步数据表明，基于CRE和FLP的 交叉法可以允许对单个ipRGC亚型进行这种操作。这项建议旨在 确定单个ipRGC亚型特有的遗传标记组合。此外，我们将同时 创建可集成到未来ipRGC亚型的任何CRE/FLP基序中的模块化遗传工具包 操纵。这些工具不仅将提供对ipRGC连接电路的即时洞察，而且将提供 视觉和回路神经科学界研究广泛的关键工具和实验管道 将细胞功能与行为联系起来的一系列电路。

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.