Richardson S B, Nguyen T, Hollander C S
Am J Physiol. 1983 Jun;244(6):E560-6. doi: 10.1152/ajpendo.1983.244.6.E560.
We studied the release of immunoreactive somatostatin (IR-SRIF) from hypothalamic cells that were obtained from rats and dispersed with the aid of collagenase. Twenty-four hours after dispersion, cells were placed in a column supported by a matrix of preswollen Biogel P2 and perifused. Fractions were collected on ice and subsequently assayed for SRIF. SRIF release was stimulated markedly by potassium depolarization (KCl, 56 mM), by the Na+-K+-ATPase inhibitor ouabain (10(-4) M), and by dopamine at concentrations as low as 10(-11) M. The stimulatory effects of membrane depolarization were calcium dependent and were not observed in the absence of exogenous calcium in the perifusion medium or in the presence of EDTA (0.05 M). Metoclopramide, the dopamine antagonist, abolished the stimulatory effect of dopamine. In conclusion, release of IR-SRIF by dispersed rat hypothalamic cells can be studied in a simple perifusion apparatus. Release is stimulated by membrane depolarization in a calcium-dependent manner and by dopamine at physiological concentrations.
我们研究了从大鼠获取并借助胶原酶分散的下丘脑细胞中免疫反应性生长抑素(IR - SRIF)的释放情况。分散24小时后,将细胞置于由预溶胀的Biogel P2基质支撑的柱中进行灌流。收集的馏分置于冰上,随后测定SRIF。钾去极化(56 mM KCl)、Na + - K + - ATP酶抑制剂哇巴因(10(-4) M)以及低至10(-11) M浓度的多巴胺均可显著刺激SRIF的释放。膜去极化的刺激作用依赖于钙,在灌流介质中无外源钙或存在EDTA(0.05 M)时未观察到该作用。多巴胺拮抗剂甲氧氯普胺消除了多巴胺的刺激作用。总之,可在简单的灌流装置中研究分散的大鼠下丘脑细胞释放IR - SRIF的情况。释放受膜去极化以钙依赖方式以及生理浓度的多巴胺刺激。