NEUROPEPTIDE Y MODULATION OF ADRENOMEDULLARY SECRETION

神经肽 Y 对肾上腺髓质分泌的调节

• 批准号: 3412351
• 负责人: TERRY D HEXUM
• 金额: $ 12.65万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1994-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3412351
• 关键词: G protein; adrenal medulla; antisense nucleic acid; binding proteins; catecholamines; chromaffin cells; hippocampus; neuropeptide Y; neuropeptide receptor; oligonucleotides; radiotracer; second messengers; secretion

The overall significance of our proposed studies is to further the understanding of the physiological relevance of peptide action in neurotransmitter function in general and on adrenomedullary secretion in particular. Pilot studies have shown that low concentrations (1 X 10-8 M) of neuropeptide Y (NPY) inhibit nicotine stimulated adrenomedullary secretion. Our hypothesis is that NPY exerts its effect on adrenomedullary catecholamine secretion by modulating nicotinic receptor stimulation. The specific aims are to : 1) delineate the effects and structural requirements of NPY on nicotinic receptor stimulation, 2) determine whether NPY in chromaffin cells influences secretion, 3) describe NPY binding and its effect on agonist binding to the nicotinic receptor, and 4) identify and characterize the role of G-binding proteins and phosphoinositide turnover in the modulatory action of NPY. The modulatory effect of NPY will be examined by measuring the nicotine receptor-induced secretion of epinephrine, norepinephrine and the enkephalin-like peptides (ME-IRM) using cultured bovine chromaffin cells. Catecholamine and ME-IRM secretion will be measured by high performance liquid chromatography with electrochemical detection, and with radioimmunochemical techniques using a specific antibody to (Met5)-enkephalin, respectively. NPY influence on secretion will be determined by studying it's action in the presence of various nicotinic agonists, it's action on agents stimulating secretion through other than nicotinic receptors and the effect of truncated forms of NPY and peptides possessing sequence homologies with NPY. The role of endogenous NPY will be examined through the use of antibodies and antisense RNA to NPY. The mechanism of the modulatory effect of NPY on secretion will be examined by studying the effect of NPY on nicotinic receptor binding and on changes produced in the second messengers (G-binding proteins, phosphoinositide turnover) that may be involved in the transduction of the message received during nicotinic receptor stimulation. We will employ the techniques of cell culture, radioimmunoassay, high performance liquid chromatography, antisense oligonucleotide probes, radioreceptor binding, ion-exchange chromatography and thin layer chromatography. The health related consequences of the proposed studies are far-reaching due to the varied roles suggested for neuropeptides in physiology and pathology, e.g., defective neuronal regulation leading to hypertension or mental disorders.

我们提出的研究的总体意义是进一步 了解肽作用的生理相关性 神经递质的一般功能和肾上腺髓质分泌 特别的。 初步研究表明，低浓度 (1 X 10-8 M) 神经肽 Y (NPY) 抑制尼古丁刺激的肾上腺髓质 分泌。 我们的假设是 NPY 对肾上腺髓质发挥作用 通过调节烟碱受体刺激来分泌儿茶酚胺。 这 具体目标是： 1) 描述效果和结构要求 NPY对烟碱受体刺激的影响，2)确定NPY是否在 嗜铬细胞影响分泌，3) 描述 NPY 结合及其 对激动剂与烟碱受体结合的影响，以及 4) 识别和 表征 G 结合蛋白和磷酸肌醇周转的作用 NPY 的调节作用。 NPY 的调节作用为 通过测量尼古丁受体诱导的分泌来检查 使用肾上腺素、去甲肾上腺素和脑啡肽样肽 (ME-IRM) 培养的牛嗜铬细胞。 儿茶酚胺和 ME-IRM 分泌会 通过高效液相色谱法和电化学分析仪进行测量 检测，并使用特定的放射免疫化学技术 分别针对 (Met5)-脑啡肽的抗体。 NPY对分泌的影响 将通过研究它在存在各种情况下的行为来确定 烟碱激动剂，它通过刺激分泌的物质起作用 除烟碱受体外以及截短形式的 NPY 和的作用 与 NPY 具有序列同源性的肽。 内源性的作用 NPY 将通过使用抗体和反义 RNA 进行检查 NPY。 NPY对分泌的调节作用的机制是 通过研究 NPY 对烟碱受体结合和对烟碱受体结合的影响来检查 第二信使（G结合蛋白， 磷酸肌醇周转）可能参与转导 在烟碱受体刺激期间收到的消息。 我们将聘用 细胞培养技术、放射免疫分析技术、高效液相技术 色谱法、反义寡核苷酸探针、放射性受体结合、 离子交换色谱法和薄层色谱法。 健康状况 拟议研究的相关后果是深远的，因为 神经肽在生理学和病理学中的不同作用，例如， 神经调节缺陷导致高血压或精神障碍。