DEVELOPMENT AND FUNCTION OF SYNAPTIC CO-TRANSMISSION

这项申请的目标是加强正在进行的研究 交感神经节和TO突触共同传递的生理学 学习将在新研究中应用的分子和生物学方法 神经元分化的结果。私家侦探是一名 生理学，并在RO1的支持下进行05年的独立研究。 有良好的校际环境和充足的支持服务 在当地大学社区内。所提议的计划将确保 继续发展私家侦探S的研究事业。这将限制增长 与研究无关的活动，并提供资源以追求新的 方法论、实验和合作。这些实验将会 利用电生理、解剖学和生物化学的结合 方法分离制备法和原代细胞培养法。新的 学习的方法包括核酸分离、Northern分析和 原位杂交。 该提案的实验目标是确定综合的 利用缓慢的突触的功能和发育起源 除了经典的多肽能协同递质之外的作用 递质乙酰胆碱(ACH)和肾上腺素(EPI)。 促黄体生成素释放激素4种神经肽的作用 LHRH、神经肽Y(NPY)、降钙素基因相关肽(CGRP)和 P物质(SP)，将在牛蛙的交感系统中进行分析。 尽管这些多肽与其他递质在许多 外周和中枢神经系统的区域，功能性 突触共同传递的动力学和个体发育在很大程度上是未知的。 牛蛙交感神经节表达多种突触机制 然而，它们可以在细胞和分子水平上进行分析 级别。在腰神经节内，交感神经有3个亚类。 神经元可以通过它们的功能来区分，电生理 特性，与脊髓和周围靶点的连通性， 并通过它们的M胆碱能受体和 神经肽。在血管运动交感C系统中，LHRH和ACH是 节前神经元共释放并诱发3种突触后电位 在共表达EPI和NPY样神经节细胞的子集中 免疫反应性。这项提案的第一个具体目标是测试 神经节和终末器官突触的共同传递假说 这个回路增强了血管收缩的动态范围，可以 被不同的节前刺激模式所诱发。这个 第二个具体目标是检验CGRP共同释放的假设 通过节前B神经元的ACH并产生营养效应和 B神经元的兴奋性突触后电位。第三个目标是 为特定细胞在蝌蚪发育过程中建立时间表 神经肽基因及其产物的表达。使用纸巾 文化，该项目的最终目标将是测试假设 神经肽Y的表达受细胞外分子调控 环境和/或细胞相互作用，这些因素还 控制血管运动神经元的电生理分化。

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.