CA++ CHANNELS AND STIMULUS SECRETION COUPLING

CA 通道和刺激分泌耦合

• 批准号: 2146218
• 负责人: CRISTINA R ARTALEJO
• 金额: $ 12.85万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2146218
• 关键词: adenosine triphosphate; animal tissue; ascorbate; calcium channel; calcium channel blockers; calcium flux; calcium indicator; cats; chromaffin cells; enkephalins; enzyme activity; epinephrine; exocytosis; neuropharmacology; norepinephrine; okadaic acid; phosphoprotein phosphatase; protein kinase A; second messengers; secretion; tissue /cell culture; vasoactive intestinal peptide; voltage /patch clamp; voltage gated channel

The overall goals of this research are to understand first, the function and regulation of chromaffin cell Ca channels and second, how Ca channels and catecholamine secretion are related and modulated in different chromaffin cells. The chromaffin cells of the adrenal medulla secrete catecholamines and several neuropeptide hormones in response to neural stimulation. Secretion is a Ca2+-dependent process that is triggered by the entry of Ca2+ through voltage-gated Ca channels in the plasma membrane. Thus, understanding the properties of Ca channels is essential to understanding the mechanism and regulation of catecholamine secretion. We have found that bovine chromaffin cells contain three distinct types of Ca channel, including a novel class known as faciliation Ca channels. The latter resemble "L-type" Ca channels found in other cells, in that they have a single channel conductance of 27 pS and are sensitive to dihydropyridines. These channels can be recruited by repetitive depolarizations in the physiological range, by depolarization to very positive potentials or by neurotransmitters that activate adenylyl cyclase and elevate intracellular cAMP by a mechanism involving PKA. Our working hypothesis is that these channels may be the underlying regulatory system in the "fight or flight" response. These cells also have two other types of Ca channels, both of which have a 14 pS conductance. One type is responsible for the "non classical" N-type Ca current, while the other carries a P-type Ca current component. We propose to: a) investigate the presence of these channels in epinephrine - and norepinephrine-containing bovine chromaffin cells as well as the feline chromaffin cells; b) study Ca channel modulation by different compounds co-stored and co-released during the exocytotic process and c) study the secretory response in the three cell types. In particular, we will try to evaluate to what extent each type of Ca channel participates in catecholamine secretion. Comparing secretion data from three different cell types with possibly different ratios of the same types of Ca channels will allow us to understand the role of such channels in the regulation of secretion. By simultaneously studying Ca currents (using patch-clamp electrophysiology techniques) and catecholamine secretion (using capacitance measurements), we hope to uncover the relationship between these two critical parameters. These studies will expand our understanding of Ca channels in general and their role in catecholamine secretion and by extension in synaptic neurotransmitter release.

本研究的总体目标是首先了解 和调节嗜铬细胞钙通道，第二，钙通道如何 和儿茶酚胺分泌相关，并在不同的调节 嗜铬细胞肾上腺髓质的嗜铬细胞分泌 儿茶酚胺类和几种神经肽激素对神经 刺激.分泌是一个依赖于Ca 2+的过程，由 钙离子通过电压门控钙通道进入血浆 膜的因此，了解钙通道的特性是至关重要的 了解儿茶酚胺分泌的机制和调节。 我们发现牛嗜铬细胞含有三种不同类型的 钙通道，包括一个新的类别称为facilization钙通道。的 后者类似于在其他细胞中发现的“L型”Ca通道，因为它们 具有27 pS的单沟道电导，并且对 二氢吡啶。这些渠道可以通过重复招募 在生理范围内的去极化，通过去极化到非常 正电位或激活腺苷酸环化酶的神经递质 并通过PKA相关机制升高细胞内cAMP。我们的工作 一种假说认为，这些渠道可能是潜在的调节系统 在“战斗或逃跑”的反应。这些细胞还有另外两种类型 的Ca通道，两者都具有14 pS的电导。一类是 负责“非经典”的N型钙电流，而另一个 携带P型Ca电流分量。我们建议：（a）调查 这些通道在含有肾上腺素和去甲肾上腺素的 牛嗜铬细胞以及猫嗜铬细胞; B）研究 不同化合物共储存和共释放对钙通道的调节 在胞吐过程中和c）研究在胞吐过程中的分泌反应。 三种细胞类型特别是，我们将尝试评估在多大程度上 每种类型的Ca通道都参与儿茶酚胺分泌。比较 来自三种不同细胞类型的分泌数据， 相同类型的钙通道的比例将使我们能够了解 这些通道在调节分泌中的作用。 通过同时研究钙电流（使用膜片钳 电生理技术）和儿茶酚胺分泌（使用 电容测量），我们希望揭示 这两个关键参数。这些研究将扩大我们对 钙通道的一般和它们在儿茶酚胺分泌中的作用， 突触神经递质释放的延长。