CELL BIOLOGY OF THE RETINA

描述（改编自申请人摘要）：本研究的总体目标 该项目将转基因技术与显微镜技术相联合收割机， 电生理学来研究小鼠视网膜的电路。 使用 转基因小鼠产生报告基因产物， 允许生理学和形态学在已知的子集中的相关性， 哺乳动物细胞 这是一项强大的技术， 目前是DA能无长突细胞。 另外两个转基因株系， 双极细胞和α神经节细胞也已被标记。 在上一个资助期间，申请人发现了一个自发的 DA能无长突细胞的爆发模式。 一个具体目标是充分 了解调节DA能无长突细胞的机制。 使用 RT-PCR，申请人计划鉴定包含在 DA能无长突细胞，然后使用电生理实验， 描述变送器电流。 超微结构免疫细胞化学 用于定位DA能无长突细胞表面的受体亚单位。 第二个具体目标是将这些方法扩展到其他神经元， 视网膜 cDNA文库将从特定的细胞类型中产生， 用于细胞鉴定的RT-PCR和报告基因。 申请人计划 在完整的视网膜或视网膜切片中表征这些神经元。 在继续发展这一具体目标的过程中，申请方计划 产生新的转基因动物品系， 神经元被标记。

DESCRIPTION (Adapted from applicant's abstract): The overall goal of this project is to combine transgenic technology with microscopy and electrophysiology to study circuitry of the mouse retina. The use of transgenic mice to generate reporter gene products that identify neurons allows a correlation of physiology and morphology in a known subset of mammalian cells. This is a powerful technique that has been applied currently to DAergic amacrine cells. Two other transgenic lines, rod bipolars and alpha ganglion cells, have also been labeled. In the previous funding period, the applicant discovered a spontaneous bursting pattern in DAergic amacrine cells. One specific aim is to fully understand the mechanisms that modulate DAergic amacrine cells. Using RT-PCR, the applicant plans to identify the receptor subunits contained in DAergic amacrine cells and then use electrophysiological experiments to describe transmitter currents. Ultrastructural immunocytochemistry will be used to localize receptor subunits on the surface of DAergic amacrine cells. The second specific aim is to expand these approaches to other neurons in the retina. cDNA libraries will be generated from specific cell types using RT-PCR and reporter genes for cell identification. The applicant plans to characterize these neurons either in the intact retina or the retinal slice. In the continuing development of this specific aim, the applicant plans to generate new lines of transgenic animals in which different populations of neurons are labeled.