FUNCTIONAL ORGANIZATION OF VERTEBRATE VISUAL SYSTEMS

这个项目的总体目标是了解视网膜 功能。在接下来的5年里，我们将集中精力尝试 识别视网膜神经元使用的神经递质和 尤其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保持的分离的视网膜细胞 持续几天到几周的时间。提出了五个具体项目：1) 隔离的水平和双极电池的贴片和电压钳制 鲤鱼和溜冰的视网膜。该项目正在通过膜片钳技术进行研究 水平细胞膜对L-谷氨酸和甘氨酸的反应 它的类似物。我们特别计划使用膜片钳电极来 电压钳位隔离的细胞，以研究发生的离子流 在这些细胞中对神经递质物质做出反应。2)分析 分离和培养细胞反应的离子机制 鲤鱼和滑冰鱼视网膜的水平和双极细胞 神经递质制剂，以及对其影响的定量评估 这些神经元上的神经递质激动剂和拮抗剂。为了这些 实验、药物或改变的林格氏溶液将应用于 通过微灌流技术分离细胞。3)生理学和 非水平培养神经元的药理学研究 双极细胞，以及一项对培养中保持的分离神经元的研究 是从鱼以外的动物中提取的。我们特别感兴趣的是 无长突细胞在培养中的维持及其生理学研究 和药理学特性，并研究分离的视网膜神经元 来自哺乳动物，也许是兔子，在培养中。4)突触形成的研究 在培养中保持的神经元和生理和 对这种突触的药理学分析。最初，我们计划研究 在培养的水平细胞之间形成电突触，但 最终我们希望诱导和研究化学突触的形成 视网膜细胞种类繁多。5)药物和药物的作用分析 改变了Ringer‘s溶液对完整的灌流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 药物和/或改良林格氏液对体外培养细胞的作用比较 对完整视网膜中的细胞具有相同处理效果的细胞。

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.