Lowy R J, Dawson D C, Ernst S A
Am J Physiol. 1985 Jul;249(1 Pt 1):C41-7. doi: 10.1152/ajpcell.1985.249.1.C41.
Primary cultures of structurally polarized sheets of avian salt gland secretory cells were mounted in Lucite chambers for transmural electrophysiological analysis. Transmural resistance values increased during the first 3 days of culture to 293 +/- 35 omega X cm2 and then decreased slowly thereafter. There was little short-circuit current (Isc) in the absence of secretagogues. Serosal addition of either carbachol or epinephrine resulted in a Isc consistent with positive charge flow from mucosa to serosa, thus demonstrating that these cell layers were capable of active ion transport in response to either cholinergic or adrenergic neurohormonal stimulation. Serosal ouabain or furosemide abolished the response to either agonist, while theophylline enhanced the response. Receptor specificity for the electrical responses was shown by selective inhibition of carbachol- and epinephrine-induced Isc by atropine and propranolol, respectively. The results demonstrate that these primary epithelial cell cultures are capable of active ion transport and are sensitive to known inhibitors of secretory transport, and suggest that intracellular coupling mechanisms for hormonal control are retained in culture. These cultures should be useful for studying mechanisms of ion secretory transport and their regulatory control.
将鸡盐腺分泌细胞结构极化片的原代培养物安装在有机玻璃小室中进行跨膜电生理分析。跨膜电阻值在培养的前3天增加到293±35Ω×cm2,此后缓慢下降。在没有促分泌剂的情况下,短路电流(Isc)很小。在浆膜侧添加卡巴胆碱或肾上腺素会导致Isc,这与正电荷从黏膜流向浆膜一致,从而表明这些细胞层能够响应胆碱能或肾上腺素能神经激素刺激进行主动离子转运。浆膜侧添加哇巴因或呋塞米会消除对任何一种激动剂的反应,而茶碱会增强这种反应。阿托品和普萘洛尔分别选择性抑制卡巴胆碱和肾上腺素诱导的Isc,显示了电反应的受体特异性。结果表明,这些原代上皮细胞培养物能够进行主动离子转运,并且对已知的分泌转运抑制剂敏感,这表明激素控制的细胞内偶联机制在培养中得以保留。这些培养物对于研究离子分泌转运机制及其调节控制应该是有用的。