Stimulus-secretion coupling in chromaffin cells isolated from bovine adrenal medulla.
Stimulus-secretion coupling in chromaffin cells isolated from bovine adrenal medulla.
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从牛肾上腺髓质分离的嗜铬细胞中的刺激-分泌耦合。
DOI：
10.1073/pnas.74.11.5036
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发表时间：
1977
影响因子：
11.1
通讯作者：
K. Rosenheck
中科院分区：
文献类型：
--
作者：
A. S. Schneider;R. Herz;K. Rosenheck
牛肾上腺嗜铬细胞通过去除皮质和顺序的胶原酶消化髓质分离。这些细胞的儿茶酚胺分泌功能的特点与乙酰胆碱刺激，阳离子的要求，和细胞骨架元素。刺激释放的剂量-反应曲线在10（-5）M乙酰胆碱时有其半最大值，最大分泌量平均为对照基础分泌量的7倍。0.1 mM乙酰胆碱刺激后，肾上腺素与去甲肾上腺素的差异释放与它们在细胞悬液中的分布成比例。胆碱能受体主要是烟碱。儿茶酚胺释放的动力学是快速的，与显着的分泌发生在不到60秒和85%的最大分泌5分钟内。一个关键的要求，在细胞外介质中的钙被证明，和80%的最大分泌是在生理钙浓度。刺激过量的钾（65 mM KCl）也诱导儿茶酚胺分泌，这不同于乙酰胆碱刺激的效力较低，在具有不同的依赖钙浓度，并在其响应的局部麻醉剂丁卡因。丁卡因，这被认为是抑制膜阳离子渗透性，能够阻止乙酰胆碱刺激，但不KCl刺激的分泌。微管破坏剂长春碱能够阻断儿茶酚胺的释放，而微丝破坏剂细胞松弛素B几乎没有影响。结果表明，分离的牛嗜铬细胞是活的，功能，并在大量的可用性。这些细胞现在为研究激素和神经递质释放的细胞表面调节提供了一个极好的系统。
Bovine adrenal chromaffin cells were isolated by removal of the cortex and sequential collagenase digestion of the medulla. The catecholamine secretory function of these cells was characterized with respect to acetylcholine stimulation, cation requirements, and cytoskeletal elements. The dose-response curve for stimulated release had its half-maximum value at 10(-5) M acetylcholine, and maximum secretion was on the average 7 times that of control basal secretion. The differential release of epinephrine versus norepinephrine after stimulation with 0.1 mM acetylcholine occurred in proportion to their distribution in the cell suspension. The cholinergic receptors were found to be predominantly nicotinic. The kinetics of catecholamine release were rapid, with significant secretion occurring in less than 60 sec and 85% of maximum secretion within 5 min. A critical requirement for calcium in the extracellular medium was demonstrated, and 80% of maximum secretion was achieved at physiologic calcium concentrations. Stimulation by excess potassium (65 mM KCl) also induced catecholamine secretion which differed from acetylcholine stimulation in being less potent, in having a different dependence on calcium concentration, and in its response to the local anesthetic tetracaine. Tetracaine, which is thought to inhibit membrane cation permeability, was able to block acetylcholine-stimulated but not KCl-stimulated secretion. The microtubule disrupting agent vinblastine was able to block catecholamine release whereas the microfilament disrupter cytochalasin B had little effect. The results show the isolated bovine chromaffin cells to be viable, functioning, and available in large quantity. These cells now provide an excellent system for studying cell surface regulation of hormone and neurotransmitter release.