FUNCTIONAL ORGANIZATION OF VERTEBRATE VISUAL SYSTEMS

这个项目的总体目标是获得视网膜的理解 功能 在接下来的五年里，我们将集中精力， 来识别视网膜神经元所使用的神经递质， 特别是这些物质对视网膜细胞的影响。 大部分 计划中的工作将利用培养中的分离视网膜细胞 持续数天至数周。 提出了五个具体项目：1） 的隔离水平和双极单元的补丁和电压钳位 鲤鱼和鳐的视网膜。 本课题采用膜片钳技术进行研究 技术水平细胞膜对L-谷氨酸的反应， 其类似物。 我们特别计划使用膜片钳电极， 电压钳分离的细胞，以研究发生的离子流 对神经递质物质的反应。 2)分析 分离的和培养的细胞的反应背后的离子机制 水平和双极细胞的鲤鱼和滑冰视网膜的各种 神经递质药物，以及定量评价的影响， 神经递质激动剂和拮抗剂。 为这些 实验，药物或改变林格氏溶液将被应用于 通过微灌注技术分离细胞。 3)一个生理和 神经元的药理学研究，而不是水平培养， 双极细胞，以及一项对培养的分离神经元的研究， 是从鱼类以外的动物身上提取的。 我们特别感兴趣 在维持无长突细胞的培养和研究其生理 以及药理学特性，同时研究分离的视网膜神经元 从哺乳动物，也许是兔子，在文化。 4)突触形成的研究 在培养的神经元和生理的， 对这种突触的药理学分析。 最初我们计划研究 培养的水平细胞之间的电突触形成，但 最终，我们希望诱导和研究化学突触的形成之间的一个 各种视网膜细胞。 5)分析了药物的作用， 改变林格氏溶液对完整灌注视网膜中神经元的影响。 这些 实验旨在通过以下方式补充分离的细胞研究： 比较药物和/或改变的林格氏溶液对分离的 细胞与对完整视网膜中的细胞的相同处理的效果。

The overall goal of this project is to gain an understanding of retinal function. For the next 5 years, we are focusing our efforts on attempting to identify the neurotransmitters used by the retinal neurons and especially on the effects these substances have on retinal cells. Much of the work planned will utilize isolated retinal cells maintained in culture for periods of days to weeks. Five specific projects are proposed: 1) Patch and voltage clamping of isolated horizontal and bipolar cells of the carp and skate retina. This project is studying by patch clamping techniques the response of the horizontal cell membrane to L-glutamate and its analogues. We plan in particular to use patch clamp electrodes to voltage clamp the isolated cells in order to study the ion flows that occur in these cells in response to neurotransmitter substances. 2) An analysis of the ionic mechanisms underlying the responses of isolated and cultured horizontal and bipolar cells of the carp and skate retinas to various neurotransmitter agents, and a quantitative evaluation of the effects of neurotransmitter agonists and antagonists on these neurons. For these experiments, drugs or altered Ringer's solution will be applied to the isolated cells by a microperfusion technique. 3) A physiological and pharmacological study of neurons in culture other than horizontal and bipolar cells, and a study of isolated neurons maintained in culture that are derived from animals other than fish. We are particularly interested in maintaining amacrine cells in culture and studying their physiological and pharmacological properties and also studying isolated retinal neurons from a mammal, perhaps rabbit, in culture. 4) A study of synapse formation between neurons maintained in culture and a physiological and pharmacological analysis of such synapses. Initially we plan to study electrical synapse formation between cultured horizontal cells, but eventually we hope to induce and study chemical synapse formation between a variety of retinal cells. 5) An analysis of the effects of drugs and altered Ringer's solution on neurons in the intact perfused retina. These experiments are intended to complement the isolated cell studies by comparing the effects of drugs and/or altered Ringer's solution on isolated cells with the effects of the same treatments on cells in the intact retina.