Catecholamine Secretion by Chemically Skinned Cultured Chromaffm Cells
Catecholamine Secretion by Chemically Skinned Cultured Chromaffm Cells
复制标题
化学剥皮培养的染色质细胞分泌儿茶酚胺
作者：
J. Brooks;S. Treml
摘要：完整嗜铬细胞的分泌系统由于质膜的选择性渗透性而不能直接进行化学操作。我们设计了一个简单的程序，化学“皮肤”（透化）培养的肾上腺髓质嗜铬细胞短暂暴露于洗涤剂皂苷。这一过程破坏了质膜的连续性，从而使我们能够绕过由细胞膜控制的分泌过程的那些方面，并使外源物质直接进入细胞分泌机制。我们在这里报告，皮肤细胞保留一个完全胜任的分泌机制，只依赖于外源性钙和MgATP。皂苷处理对细胞总儿茶酚胺含量无明显影响。分泌可以启动由MgATP或钙，只要其他是存在于介质中。去皮细胞释放的儿茶酚胺和多巴胺-β-羟化酶依赖于培养基的钙浓度。分泌的儿茶酚胺和酶的比例与细胞的相似，表明分泌发生的胞吐机制。约一半的细胞质酶乳酸脱氢酶的总细胞内容物在透化过程中释放，随后的孵育，表明质膜渗透性大分子的蛋白质。钙诱导的分泌不受已知影响儿茶酚胺和颗粒功能的几种药物的影响。在培养中的嗜铬细胞的皂苷治疗似乎是一种简单的手段，允许外源性物质进入细胞的分泌机制。因此，它提供了使用化学处理和可能的细胞成分特异性抗体的机会，以确定这些元素在分泌过程中的作用。这些技术也应该适用于已知通过胞吐机制分泌的其他细胞。
Abstract: The secretory system of intact chromaffin cells is not accessible to direct chemical manipulation because of the selective permeability of the plasmalemma. We have devised a simple procedure for chemically “skinning” (permeabilizing) cultured adrenal medullary chromaffin cells by brief exposure to the detergent saponin. This procedure disrupts the continuity of the plasmalemma, thus allowing us to bypass those aspects of the secretory process controlled by the cell membrane and giving direct access of exogenous substances to the cellular secretory machinery. We report here that the skinned cells retain a fully competent secretory mechanism dependent only on exogenous calcium and MgATP. Saponin treatment had no significant effect on the total catecholamine content of the cells. Secretion could be initiated by either MgATP or calcium as long as the other was present in the medium. Catecholamine and dopamine‐β‐hydroxylase release by the skinned cells was dependent on the calcium concentration of the medium. The ratio of secreted catecholamine and enzyme was similar to that of the cells, indicating that secretion occurred by an exocytotic mechanism. About half the total cellular content of the cytoplasmic enzyme lactic dehydrogenase was released during the permeabilization process and subsequent incubations, indicating plasmalemma permeability to molecules as large as protein. Calcium‐induced secretion was unaffected by several drugs known to affect catecholamines and granule function. Saponin treatment of chromaffin cells in culture appears to be a simple means for allowing access of exogenous substances to the cell's secretory machinery. Therefore, it offers the opportunity to use chemical treatments and perhaps specific antibodies to cellular components, to determine the role of these elements in the secretory process. These techniques should also be applicable to other cells known to secrete by an exocytotic mechanism.
完整仓鼠胰岛素瘤细胞中 Ca2 依赖性蛋白磷酸化和胰岛素释放。
DOI：
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发表时间：
1980
期刊：
The Journal of biological chemistry
影响因子：
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作者：
Schubart,UK;Fleischer,N;Erlichman,J
通讯作者：
Erlichman,J