RELEASE, STORAGE AND UPTAKE OF CATECHOLAMINES BY A CLONAL CELL LINE OF NERVE GROWTH-FACTOR (NGF) RESPONSIVE PHEOCHROMOCYTOMA CELLS
RELEASE, STORAGE AND UPTAKE OF CATECHOLAMINES BY A CLONAL CELL LINE OF NERVE GROWTH-FACTOR (NGF) RESPONSIVE PHEOCHROMOCYTOMA CELLS
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DOI：
10.1016/0006-8993(77)90005-1
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发表时间：
1977-01-01
期刊：
影响因子：
2.9
通讯作者：
REIN, G
中科院分区：
文献类型：
--
作者：
GREENE, LA;REIN, G
在指定为 PC12 的大鼠嗜铬细胞瘤细胞克隆系的培养物中研究了儿茶酚胺的释放、储存和摄取。先前显示 PC12 系通过延长神经元样过程来响应 NGF，并合成并含有去甲肾上腺素 (NE) 和多巴胺 (DA)。在本实验中，在暴露于51.5mM K+时，PC12细胞释放相当大比例(对于NGF未处理和NGF处理的细胞分别约为30和12)的内源NE和DA。该释放取决于 Ca2+ 的存在，并被过量的 Mg2+ 抑制。 Veratridine 是一种激活 Na+ 动作电位离子载体的生物碱，也能有效释放 DA 和 NE。 PC12 细胞（NGF 处理和未处理）暴露于利血平 (10-5 M) 导致细胞内 DA 和 NE 水平在 21 小时内消耗超过 90%。 PC12细胞还通过可饱和转运机制从外部介质摄取NE，该机制遵循米氏动力学(表观Km约2μM)，是能量和Na依赖性的并且被低浓度的可卡因和去甲基丙咪嗪阻断。这些发现表明，PC12细胞可以通过胞吐分泌机制释放儿茶酚胺，将大部分内源性儿茶酚胺储存在嗜铬颗粒和/或囊泡中，并具有摄取1型NE转运系统。 PC12 细胞似乎表达交感神经元和肾上腺嗜铬细胞的许多分化特性，并且可能是研究儿茶酚胺代谢的有用系统。
Release, storage and uptake of catecholamines were studied in cultures of a clonal line of rat pheochromocytoma cells designated as PC12. The PC12 line was previously shown to respond to NGF by extending neuronal-like processes and to synthesize and contain norepinephrine (NE) and dopamine (DA). In the present experiments, on exposure to 51.5 mM K+, PC12 cells released a substantial proportion (.apprx. 30 and 12 for NGF-untreated and -treated cells, respectively) of their endogenous NE and DA. The release was dependent on the presence of Ca2+ and was inhibited by excess Mg2+. Veratridine, an alkaloid which activates Na+ action potential ionophores, was also effective in releasing DA and NE. Eposure of PC12 cells (NGF treated and -untreated) to reserpine (10-5 M) resulted in depletion of intracellulr DA and NE levels by over 90% in 21 h. PC12 cells also took up NE from the external medium by a saturable transport mechanism which follows Michaelis-Menten kinetics (apparent Km .apprx. 2 .mu.M), is energy and Na dependent and is blocked by low concentrations of cocaine and desmethylimipramine. Such findings indicate that PC12 cells can release catecholamines by an exocytotic secretion mechanism, store most of their endogenous catecholamines in chromaffin granules and/or vesicles and have an uptake1 type transport system for NE. PC12 cells appear to express a number of differentiated properties characteristic of sympathetic neurons and adrenal chromaffin cells and may be a useful system in which to study catecholamine metabolism.