PHARMACOLOGY AND PHYSIOLOGY OF RETINAL GANGLION CELLS

视网膜神经节细胞的药理学和生理学

基本信息

批准号：
3465743
负责人：
ANDREW T ISHIDA
金额：
$ 7.1万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
1989
资助国家：
美国
起止时间：
1989-04-01 至 1994-03-31
项目状态：
已结题

项目摘要

The experiments described in this proposal are designed to investigate the pharmacological and physiological properties of ganglion cells of adult goldfish retinas. The specific aims of this project are to identify the types of neurotransmitter receptors present in ganglion cell membranes, and to analyze the mechanisms by which these neurotransmitters affect the electrical activity of ganglion cells. To circumvent the difficulties of measuring these effects in situ, single ganglion cells will be dissociated from freshly-dissected retinas by standard methods, and maintained until use in primary cell culture. The response of these cells to neurotransmitters and related pharmacologic agents will then be studied under voltage-clamp to avoid the limitations of extracellular and intracellular voltage recording. To minimize cell damage, to control intracellular ionic constituents, and to directly observe pharmacological effects on single-channel currents, patch-clamp methods of voltage-clamping will be used. Ganglion cells will be identified by markers transported retrogradely via the optic nerve or on the basis of morphological features recognized as unique to ganglion cells in separate retrograde-filling experiments. Ganglion cells from goldfish will be used in these studies, because most of the transmitters to be tested have been localized by histochemical and immunocytochemical methods to cells which form synaptic inputs onto these cells in situ -- particularly neuropeptides, acetylcholine, and various amino acids. This will allow us to formulate models of specific synaptic interactions in the retina by correlating the results of the electrophysiological studies described above with anatomical and histochemical studies of the pre-synaptic inputs to ganglion cells in the intact retina. These studies should provide information about the synaptic physiology and pharmacology of ganglion cells, which in turn may be useful in assessing the effects of pharmacologic agents on various visual functions, such as detection of moving stimuli and of concentrically-arranged stimuli.

本提案中描述的实验旨在研究的药理和生理特性成年金鱼视网膜的神经节细胞。具体目的该项目是确定神经递质的类型神经节细胞膜中存在的受体，并分析这些神经递质影响电的机制神经节细胞的活性。避免困难测量原位这些影响，单个神经节细胞将是通过标准方法与新鲜删除的视网膜分离，并且保持直到原代细胞培养为止。反应这些细胞到神经递质和相关的药理剂然后将在电压钳下研究以避免限制细胞外和细胞内电压记录。最小化细胞损伤，以控制细胞内离子成分，然后直接观察到对单通道的药理作用电流，贴片钳方法的电压钳方法将使用。神经节细胞将通过运输的标记来识别通过视神经或形态学逆转在单独的神经节细胞中识别为独特的特征逆行填充实验。金鱼的神经节细胞将在这些研究中使用，因为大多数发射机是测试已通过组织化学和免疫细胞化学定位在这些单元上形成突触输入的细胞的方法原位 - 尤其是神经肽，乙酰胆碱和各种氨基酸。这将使我们能够制定特定的模型通过将视网膜突触相互作用通过关联上面描述的电生理研究和神经节前输入的组织化学研究完整视网膜中的细胞。这些研究应提供有关突触生理和药理学的信息神经节细胞又可能有助于评估药理学对各种视觉功能的影响，作为移动刺激的检测和浓度的刺激。