Okada Y, Ueda S
J Physiol. 1984 Sep;354:109-19. doi: 10.1113/jphysiol.1984.sp015366.
Fragments of the gastric fundus of 6-8-day-old rats were maintained in tissue culture. From the explant, adhered to a plastic substrate, epithelial cells migrated and developed to form a monolayer colony. Histological and histochemical studies as well as indirect immunofluorescence studies using anti-parietal cell antibodies testified to the presence of parietal cells in the monolayer during the first week. These parietal cells were distinguished by their vesicular cytoplasmic structures using phase-contrast or differential interference-contrast microscopy. Acridine Orange, an optical probe of H+ accumulation, was taken up preferentially by these parietal cells, exhibiting orange fluorescence within the cells on the third day of culture, in response to stimulation with gastrin, histamine and carbachol. The resting potential of these cultured parietal cells was about -20 mV. On day 2-4 of culture, the cell membrane became hyperpolarized (up to -30 to -40 mV) in response to gastrin, carbachol or histamine in the presence of isobutylmethyl-xanthine (IMX). During hyperpolarization, the membrane resistances decreased significantly. The amplitude and the polarity of secretagogue-induced responses were found to be dependent on the extracellular concentration of K+ (but not Na+ and Cl-). The carbachol-induced responses were inhibited by atropine but not curare. The responses induced by histamine plus IMX were blocked by cimetidine but not pyrilamine. Neither atropine nor cimetidine affected the gastrin-evoked responses. It is concluded that rat parietal cells have separate receptors for acetylcholine (muscarinic), gastrin and histamine (H2), and that an increase in the membrane permeability to K+ is closely associated with the responses of these receptors under these in vitro conditions.
将6 - 8日龄大鼠的胃底碎片进行组织培养。从附着于塑料基质的外植体上，上皮细胞迁移并发育形成单层集落。组织学、组织化学研究以及使用抗壁细胞抗体的间接免疫荧光研究证实，在第一周单层中存在壁细胞。使用相差显微镜或微分干涉对比显微镜观察，这些壁细胞以其泡状细胞质结构为特征。吖啶橙是一种H⁺积累的光学探针，在培养第三天，受到胃泌素、组胺和卡巴胆碱刺激后，这些壁细胞优先摄取吖啶橙，细胞内呈现橙色荧光。这些培养的壁细胞的静息电位约为 - 20 mV。在培养的第2 - 4天，在异丁基甲基黄嘌呤（IMX）存在的情况下，细胞膜对胃泌素、卡巴胆碱或组胺发生超极化（高达 - 30至 - 40 mV）。在超极化期间，膜电阻显著降低。发现促分泌剂诱导反应的幅度和极性取决于细胞外K⁺浓度（而非Na⁺和Cl⁻）。卡巴胆碱诱导的反应被阿托品抑制，但不被箭毒抑制。组胺加IMX诱导的反应被西咪替丁阻断，但不被吡苄明阻断。阿托品和西咪替丁均不影响胃泌素诱发的反应。结论是大鼠壁细胞具有乙酰胆碱（毒蕈碱型）、胃泌素和组胺（H2）的独立受体，并且在这些体外条件下，膜对K⁺通透性的增加与这些受体的反应密切相关。
