CELL BIOLOGY OF THE RETINA

这个研究项目的目的是了解视网膜 分析视觉世界，并编码其空间，时间 和色彩对比度转化为动作电位的信息， 发送到大脑。 建议调查哪种化学品 信息汇聚在每种类型的视网膜神经元上， 这些信息，它们在细胞表面的目的地， 混元 此外，如果能够在体外识别， 视网膜分离，神经元代表这种目标 信使分子，人们可以将它的布线与细胞的 physiology. 一个人还将拥有一个战略， 生理实验和调查的影响， 配体上的神经活性物质的适当星座-和 突触后细胞的电压门控电流。 有了这些 结合结构和功能的知识，如果一个人可以识别 在形态学上，各种受体和离子通道 每种类型的视网膜神经元表达，人们也许可以预测 神经相互作用的性质，在网站上是无法访问的， 生理实验 本申请的目的，除了 完成正在进行的关于兔子双极细胞的工作， 将联合收割机分子生物学与显微镜和电生理学相结合， 研究小鼠视网膜的功能性线路。 均匀 视网膜神经元的群体将通过引入 小鼠基因组嵌合构建体，其由报告基因和 其产物参与视觉处理的基因的启动子。 的 报告基因是（i）碱性磷酸酶，其可以在两个位点检测到， 光学和电子显微镜，技术简单可靠;（ii） β-半乳糖苷酶，可以在活细胞中鉴定， 荧光染料。 启动子将是那些调节转录的启动子 编码多肽的基因， 代谢、受体和离子通道。 的形态参数 将研究标记的细胞群及其突触 连接. 此外，携带报告基因的活细胞将 在视网膜分离后在体外进行鉴定：这样， 将有可能研究的电压和配体门控电流的 通过全细胞的方式携带转基因的视网膜神经元 膜片钳技术，并将由此获得的数据整合到 神经网络是解剖学上描述的。 这些研究 是理解和合理对待 视网膜和神经系统的疾病。

The objective of this research project is to understand how the retina of mammals analyzes the visual world and encodes its spatial, temporal and chromatic contrast into a message of action potentials for safe sending to the brain. It is proposed to investigate which chemical messages are converging upon each type of retinal neuron, the weight of these messages, their destination at the cell surface and their neuron of origin. Furthermore, if one were able to recognize in vitro, after retinal dissociation, the neuron that represents the target of such messenger molecules, one could correlate its wiring with the cell's physiology. One would also possess a strategy for designing critical physiological experiments and investigate the influence of the appropriate constellation of neuroactive substances on the ligand- and voltage-gated currents of the postsynaptic cell. Armed with this combined knowledge of structure and function, if one could identify morphologically the various species of receptors and ion channels expressed by each type of retinal neuron, one could perhaps predict the nature of the neural interactions at sites that are not accessible to physiological experimentation. Aim of this application, in addition to that of completing ongoing work on the bipolar cells of the rabbit, is to combine molecular biology with microscopy and electrophysiology in the study of the functional wiring of the mouse retina. Homogeneous populations of retinal neurons will be labeled by introducing into the mouse genome chimeric constructs consisting of a reporter gene and the promoters of genes whose products participate in visual processing. The reporter genes are (i) alkaline phosphatase that can be detected at both light and electron microscopes with simple and reliable techniques; (ii) beta-galactosidase that can be identified in living cells with a fluorescent dye. The promoters will be those that regulate transcription of genes coding for peptides, rate-limiting enzymes of transmitter metabolism, receptors and ion channels. The morphological parameters of the labeled cell populations will be studied as well as their synaptic connections. Furthermore, living cells that carry the reporter gene will be identified in vitro after dissociation of the retina: in this way, it will be possible to study the voltage- and ligand-gated currents of the retinal neurons that carry the transgene by means of the whole-cell patch clamp technique and integrate the data thus obtained into the neural networks that were described anatomically. These sort of studies are fundamental to the understanding and rational treatment of the disorders of the retina and nervous system in general.