Action potential generation in reaggregates of rat aortic smooth muscle cells in primary culture.
Action potential generation in reaggregates of rat aortic smooth muscle cells in primary culture.
复制标题
原代培养中大鼠主动脉平滑肌细胞重新聚集的动作电位产生。
DOI：
10.1159/000158206
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发表时间：
1979
期刊：
影响因子：
--
通讯作者：
N. Sperelakis
中科院分区：
文献类型：
--
作者：
D. Harder;N. Sperelakis
以成年大鼠主动脉为材料制备血管平滑肌细胞。通过胶原酶/弹性酶将细胞分散，并将其镀在玻璃纸上，使其重新聚集成直径为50-200微米的小球体。这些原代培养物孵育5-14天，然后用微电极刺穿。平均静息电位为-55 mV，平均输入电阻为9.0 M ω。细胞在电和机械上都是静止的，电刺激通常不会引起反应。然而，添加ba++ (1mm)或四乙基铵离子（TEA; 5- 15mm）诱导兴奋性（伴随收缩），要么作为自发动作电位，要么通过允许对电刺激的反应。这些试剂使细胞部分去极化（例如，至- 36mv），并且输入电阻增加。与预期的起搏器行为一样，极化电流脉冲影响了ba++诱导的尖峰的自发放电频率。洗浴液中Ca++的升高增加了动作电位的幅度（超调），维拉帕米（10(-5)M）阻断了动作电位的峰值。电镜显示，重聚集体由细长的小直径细胞紧密堆积而成，其中一些细胞表现出粗细的肌丝和“致密体”；许多细胞表面有小泡。本研究结果表明，原代培养中动脉平滑肌的重组可以维持完整动脉平滑肌的功能和形态特征，为血管平滑肌功能和控制的研究提供了有益的准备。
Cultured vascular smooth muscle cells were prepared from adult rat aortas. The cells were dispersed by collagenase/elastase, and allowed to reaggregate into small spheres (50-200 micron in diameter) by plating on to cellophane. These primary cultures were incubated for 5-14 days, and then impaled with microelectrodes. The mean resting potential was -55 mV, and the mean input resistance was 9.0 M omega. The cells were quiescent, electrically and mechanically, and electrical stimulation usually did not elicit responses. However, addition of Ba++ (1 mM) or tetraehtylammonium ion (TEA; 5-15 mM) induced excitability (with accompanying contractions), either as spontaneous action potentials or by allowing responses to electrical stimulation. The cells became partially depolarized (e.g., to -36 mV) by these agents, and the input resistance increased. The frequency of spontaneous firing of the Ba++-induced spikes was affected by polarizing current pulses, as expected for pacemaker behavior. Elevation of Ca++ in the bathing solution increased the amplitude (overshoot) of the action potentials, and the spikes were blocked by verapamil (10(-5) M). Electron microscopy showed that the reaggregates consisted of a tight packing of elongated small-diameter cells, some of which exhibited thick and thin myofilaments and 'dense bodies'; many cells possessed surface caveolae. The results of this study demonstrate that reaggregates of arterial smooth muscle in primary culture can maintain functional and morphological characteristics of intact arterial smooth muscle, and therefore provide a useful preparation for the study of vascular smooth muscle function and control.