DEVELOPMENT AND FUNCTION OF SYNAPTIC CO-TRANSMISSION

突触协同传递的发育和功能

• 批准号: 3075151
• 负责人: JOHN P HORN
• 金额: $ 7.18万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-12-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3075151
• 关键词: RNA; Rana; acetylcholine; calcitonin gene related peptide; cell differentiation; electrophoresis; electrophysiology; epinephrine; gene expression; in situ hybridization; neural transmission; neurogenesis; neuropeptide Y; neuropeptide receptor; neuropeptides; neurotransmitter metabolism; nucleic acid hybridization; substance P; sympathetic ganglion; synaptogenesis; tissue /cell culture

The objectives of this application are to intensify ongoing research on the physiology of synaptic cotransmission in sympathetic ganglia and to learn molecular and biological methods that will be applied in new studies of neuronal differentiation. The P.I. is an assistant professor of Physiology and is in the 05 year of independent research with RO1 support. An excellent collegial environment and adequate support services exist within the local University community. The propose plan will insure continued development of the P.I.'s research career. It will limit growth of non-research related activities and provide resources to pursue new methodologies, experiments and collaborations. The experiments will utilize a combination of electrophysiological, anatomical and biochemical methods in isolated preparations and in primary cell cultures. The new methods to be learned include nucleic acid isolation,Northern analysis and in situ hybridization. The experimental goals of the proposal are to determine the integrative function and the developmental origins of synapses that utilize slowly acting peptidergic co-transmitters in addition to the classical transmitters acetylcholine (ACH) and epinephrine (EPI). The roles of 4 neuropeptides, luteinizing hormone releasing hormone (LHRH), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP) and substance P (SP), will be analyzed in the sympathetic system of bullfrog. Although these peptides are co-expressed with other transmitters in many regions of the peripheral and central nervous system, the functional dynamics and ontogeny of synaptic co-transmission is largely unknown. Bullfrog sympathetic ganglia express a diversity of synaptic mechanisms and yet they are accessible to analysis at the cellular and molecular levels. In the lumbar ganglia, there exist 3 subclasses of sympathetic neurons that can be distinguished by their functions, electrophysiological properties, connectivity with the spinal cord and with peripheral targets, and by their expression of muscarinic cholinergic receptors and neuropeptides. In the vasomotor sympathetic C system, LHRH and ACH are co-released by preganglionic neurons and elicit 3 postsynaptic potentials in a subset of ganglion cells that co-express EPI and NPY-like immunoreactivity. The first specific aim of this proposal is to test the hypothesis that co-transmission at ganglionic and end-organ synapses in this circuit enhances the dynamic range of vascular contractions that can be elicited by different patterns of preganglionic stimulation. The second specific aim is to test the hypothesis that CGRP is co-released with ACH by preganglionic B neurons and produces trophic effects and an excitatory postsynaptic potential in B neurons. The third goal is to establish the timetable during development in tadpoles for cell-specific expression of neuropeptide genes and their products. Using tissue culture, the final goal of the project will be to test the hypothesis that expression of NPY is controlled by molecules in the extracellular environment and/or by cellular interactions, and that such factors also control the electrophysiological differentiation of vasomotor neurons.

该应用程序的目标是加强对 同情神经节中突触共透析的生理学 学习将在新研究中应用的分子和生物学方法 神经元分化。 P.I.是 生理学，并在独立研究的05年中获得RO1支持。 存在一个极好的大学环境和足够的支持服务 在当地大学社区。 建议的计划将确保 继续发展P.I.研究生涯。它将限制增长 非研究相关的活动，并提供追求新的资源 方法，实验和协作。 实验会 利用电生理，解剖学和生化的结合 孤立制剂和原代细胞培养物中的方法。 新的 要学习的方法包括核酸隔离，北部分析和 原位杂交。 该提案的实验目标是确定综合性 慢慢使用的功能和突触的发展起源 除经典外 发射机乙酰胆碱（ACH）和肾上腺素（EPI）。 4种神经肽的作用，黄体生成激素释放激素 （LHRH），神经肽Y（NPY），降钙素基因相关肽（CGRP）和 物质P（SP）将在牛蛙的交感神经系统中进行分析。 尽管这些肽与许多其他发射机共表达 外围和中枢神经系统的区域，功能 突触共传递的动力和个体发育在很大程度上是未知的。 牛蛙的同情神经节表达了多种突触机制 但是它们可以在细胞和分子上进行分析 水平。 在腰神经节中，有3个同情的子类 可以通过其功能（电生理学）区分的神经元 特性，与脊髓的连通性以及与周围目标的连通性， 并通过表达毒蕈碱胆碱能受体和 神经肽。 在血管舒缩交感C系统中，LHRH和ACH是 通过前神经神经元共同发行并引起3突触后电位 在共表达EPI和NPY样细胞的子集中 免疫反应性。 该建议的第一个具体目的是测试 假设在神经节和末端器官突触中共转移 该电路增强了可动的血管收缩的动态范围 通过不同模式的前神经节刺激引起。 这 第二个具体目的是检验CGRP共同发布的假设 通过ganglionic B神经元进行ACH并产生营养作用，A B神经元中的兴奋性突触后潜力。 第三个目标是 在t的开发过程中建立时间表以特异性细胞特异性 神经肽基因及其产物的表达。 使用组织 文化，该项目的最终目标是检验以下假设 NPY的表达受细胞外分子的控制 环境和/或通过细胞相互作用，此类因素也 控制血管舒缩神经元的电生理分化。