CELL COMMUNICATION IN RETINA

视网膜中的细胞通讯

• 批准号: 6178592
• 负责人: RAMON F DACHEUX
• 金额: $ 34.72万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6178592
• 关键词: cell cell interaction; cone cell; electron microscopy; evoked potentials; gamma aminobutyrate; glycine; immunochemistry; laboratory rabbit; light intensity; neural information processing; neuroanatomy; neurophysiology; neurotransmitters; retina; retinal bipolar neuron; retinal ganglion; rod cell; synapses; tissue /cell culture; visual phototransduction; voltage /patch clamp

DESCRIPTION (Adapted From The Applicant's Abstract): This study is directed at understanding the morphological and physiological organization of the inner plexiform layer of the mammalian retina, and focuses on examining the synaptology related to physiologically identified ganglion cells in the rabbit retina. The present study intends to characterize the transmitter and receptors in the context of the microcircuits responsible for a specific physiological response. Direct information pertaining to the neurochemical characteristics of the synaptic profiles will be obtained by combining intracellularly HRP filled cell with electron microscopic post-embedding immunogold-cytochemistry. An analysis of transmitters and their receptor subtypes involved in microcircuits impinging on the direction selective (DS) and alpha ganglion cells will then be made. The DS and alpha ganglion cells will be injected with HRP, reacted to recover the stained cell, prepared for electron microscopy and embedded in plastic. The stained cell will be serially thin sectioned and two different antibodies will be used to label alternate sections from the continuous series. The GABA and ChAT antibodies will be used with the DS cell; GABA and glycine antibodies will be used on the alpha cell. The HRP stained dendrites with their associated synaptic profiles will be examined with electron microscopy, photographed, and printed to form a montage encompassing all the processes related to the HRP stained dendrite. The montage for each section containing its synaptic profiles will be traced into the computer, and the GABA, ChAT and glycine immunoreactive processes will be color coded. By 3-dimensionally reconstructing each synaptic profile, the serial sections containing immunoreactive positive processes will be superimposed, which allows the double labeled processes to be identified and demonstrate the synaptic interactions between differently labeled processes. In addition, the antibodies to the a- and b-subunits for the GABAA receptor and the a- and b-subunits for the acetylcholine receptor will be used to localize and determine the specific synaptology involved in the synaptic interactions between the cholinergic, starburst amacrine cell and the DS ganglion cell as well as between two cholinergic cells. Whole-cell patch-clamp recordings will be made from cone bipolar cells and alpha ganglion cells to correlate the immunocytochemical results to the presence or absence of GABA- and glycine-gated currents on these cells.

描述（根据申请人的摘要改编）：该研究针对 了解内在的形态学和生理组织 哺乳动物视网膜的丛状层，专注于检查 与生理鉴定的兔子神经节细胞有关的突触学 视网膜。本研究旨在表征发射器和受体 在负责特定生理的微电路的背景下 回复。与神经化学特征有关的直接信息 突触轮廓将通过将细胞内HRP填充来获得 带有电子显微镜后填充免疫元素化学化学的细胞。一个 分析发射器及其受体亚型参与微电路 撞击方向选择性（DS）和α神经节细胞将是 制成。 DS和Alpha神经节细胞将注射HRP，对 恢复染色的细胞，制备用于电子显微镜并嵌入 塑料。染色的细胞将串行薄截面，并且两个不同 抗体将用于标记连续系列的替代部分。 GABA和聊天抗体将与DS细胞一起使用； GABA和甘氨酸 抗体将用于α细胞上。 HRP染色的树突与他们 相关的突触轮廓将通过电子显微镜检查， 拍摄并打印以形成一个包含所有过程的蒙太奇 与HRP染色的树突有关。每个部分包含的蒙太奇 它的突触配置文件将被追溯到计算机，GABA，聊天和 甘氨酸免疫反应过程将进行颜色编码。 通过3维重建每个突触轮廓，串行部分 包含免疫反应性阳性过程将被叠加，这允许 要识别的双重标记过程并演示突触 不同标记的过程之间的相互作用。另外，抗体 到GABAA受体的A-和B-Subunits以及A-和B-Subunits的 乙酰胆碱受体将用于定位和确定特定 参与胆碱能之间突触相互作用的突触学 Starburst amacrine细胞和DS神经节细胞以及两个之间 胆碱能细胞。 全细胞斑块钳记录将由锥双极细胞和 α神经节细胞将免疫细胞化学结果与 这些细胞上存在或不存在GABA和甘氨酸门控电流。