NEURONAL NICOTINIC ACETYLCHOLINE RECEPTORS

神经元烟碱乙酰胆碱受体

• 批准号: 3408524
• 负责人: PETER B. SARGENT
• 金额: $ 8.78万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-03-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3408524
• 关键词: Anura; acetylcholine; antigens; autonomic ganglion; autoradiography; bungarotoxins; chemical binding; denervation; developmental neurobiology; electron microscopy; fresh water environment; immunochemistry; innervation; membrane activity; monoclonal antibody; neural information processing; neural transmission; nicotinic receptors; striated muscles; superior colliculus; synapses

The long-term objective of this research is to characterize neuronal nicotinic acetylcholine receptors (nAChRs) as regards to their antigenic structure, their distribution on the neuronal surface and the control of their expression during development. Neuronal nAChRs in the optic tectum and in autonomic ganglia of the frog will be studied using cross-reacting monoclonal antibodies (mAbs) made against purified nAChRs from electric organ and skeletal muscle and provided by Dr. Jon Lindstrom. In preliminary results a number of these mAbs have been found to bind to that part of the optic tectum of Rana pipiens which receives a major projection from the retina. These results are interesting in light of previous work indicating that Alpha-bungarotoxin and other nicotinic antagonists appear to block retinotectal transmission in several lower vertebrate species. The first specific aim will be to further characterize the molecule(s) in the tectum that are recognized by anti-receptor mAbs. If these mAbs indeed recognize receptors which subserve retinotectal synaptic transmission, they should bind to the surface of tectal neurons immediately beneath ganglion cell terminals. This prediction will be tested at the fine structural level using immunoperoxidase and immunogold techniques while simultaneously labeling ganglion cell afferents to the tectum. The binding of Alpha-bungarotoxin will also be visualized using HRP-labeled Alpha-bungarotoxin, and the sites to which toxin binds will be compared to mAb binding sites. The second specific aim will be to use cross-reacting polyclonal and monoclonal antibodies to characterize nAChRs on the surface of autonomic ganglion cells. Antibodies which bind to the postsynaptic membrane will be assumed to cross-react with nAChRs on the ganglion cell surface. The distribution of ganglionic nAChRs will be measured using immunogold techniques or electron microscopic autoradiography to measure the extent to which neuronal nAChRs are concentrated in the subsynaptic membrane. The influence of the presynaptic nerve upon nAChR distribution will be determined by studying antibody binding before and after preganglionc denervation. These studies will serve to improve our understanding of the structure and regulation of neuronal acetylcholine receptors.

这项研究的长期目标是表征神经元 烟碱乙酰胆碱受体（NACHRS）关于其抗原 结构，它们在神经元表面的分布以及控制 它们在发展过程中的表达。 视膜中的神经元NACHRS 在青蛙的自主神经节中，将使用交叉反应研究 针对电纯化的NACHR制成的单克隆抗体（mAb） 器官和骨骼肌，由Jon Lindstrom博士提供。 在 初步结果已经发现许多这些mAb与之结合 Rana Pipiens的光学层的一部分，它接收了主要投影 来自视网膜。 鉴于先前的工作，这些结果很有趣 表明Alpha-Bungarotoxin和其他烟碱拮抗剂出现 阻止几种下脊椎动物物种中的视网膜直肠传播。 第一个具体目的是进一步表征分子 由抗受体mAb识别的底座。 如果这些mab确实 识别能够得到视网膜直直直骨传播的受体，它们 应该与紧邻神经节下的张直肠神经元的表面结合 细胞末端。 该预测将在精细的结构上进行测试 使用免疫过氧化物酶和免疫金技术的水平同时进行 将神经节细胞传入标记为tectum。 结合 Alpha-bungarotoxin也将使用HRP标签可视化 α-bungarotoxin，以及将毒素结合的位点与 mAb结合位点。 第二个特定目的是使用交叉反应多克隆和 单克隆抗体表征在自主神经表面的NACHR 神经节细胞。 与突触后膜结合的抗体将是 假定在神经节细胞表面上与NACHR交叉反应。 这 神经节NACHR的分布将使用免疫金测量 技术或电子显微自显影术，以测量 哪些神经元NACHR集中在突触膜上。 这 突触前神经对NACHR分布的影响将是 通过研究抗体结合前后的抗体结合来确定 神经。 这些研究将有助于提高我们对结构和 神经元乙酰胆碱受体的调节。