CELL BIOLOGY OF THE RETINA

这项研究项目的目的是了解视网膜是如何 分析视觉世界并编码其空间、时间 和色度对比为安全的动作电位信息 发送到大脑。有人建议调查哪种化学物质 信息正在汇聚到每种类型的视网膜神经元上，重量 这些信息，它们在细胞表面的目的地和它们的神经元 原产地。此外，如果一个人能够在体外识别，在 视网膜解离，代表这种情况的目标的神经元 信使分子，人们可以将它的线路与细胞的 生理学。一个人还将拥有一种设计关键 生理实验，并考察了不同浓度的生理活性物质对 配体上适当的神经活性物质星座-和 突触后细胞的电压门控电流。用这个武装起来 结合了结构和功能的知识，如果一个人能识别 在形态上，各种受体和离子通道 由每种类型的视网膜神经元表达，人们可能可以预测 无法访问的位置上的神经相互作用的性质 生理实验。此应用程序的目的除了 完成正在进行的关于兔子双极细胞的工作，是 将分子生物学与显微镜和电生理学相结合 小鼠视网膜功能连接的研究。同质 视网膜神经元的种群将通过引入到 小鼠基因组嵌合结构由报告基因和 基因的启动子，其产物参与视觉处理。这个 报告基因是(I)在两个位点都能检测到的碱性磷酸酶 具有简单可靠技术的光学显微镜和电子显微镜； 可在活细胞中鉴定的β-半乳糖苷酶 荧光染料。启动子是那些调节转录的启动子 编码多肽的基因，递质的限速酶 代谢、受体和离子通道。海藻的形态参数 将研究标记的细胞群以及它们的突触 关系。此外，携带报告基因的活细胞将 在视网膜解离后被体外鉴定：通过这种方式， 将有可能研究电压门控和配体门控电流。 通过全细胞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集成到 从解剖学上描述的神经网络。这类研究 对于理解和合理对待 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.