Suzuki M, Ikeda K, Sunose H, Hozawa K, Kusakari C, Katori Y, Takasaka T
Department of Otolaryngology, Tohoku University School of Medicine, Sendai, Japan.
Hear Res. 1995 Jun;86(1-2):68-76. doi: 10.1016/0378-5955(95)00055-9.
The primary culture of marginal cells from the explant of the stria vascularis of guinea-pigs was established and confirmed by morphological, immunohistochemical and physiological characters of the cultivated cells. The positive stain of cytokeratin 18 without vimentin and desmin expressions indicated the epithelial origin of the cultured polygonal cells. Electron microscopical findings of cultured cells resembled the morphological characteristics of marginal cells. The addition of 100 microM ouabain to the cultured cells significantly increased the intracellular Na+ concentration, providing the evidence for the presence of Na+ pump. Using the cultured marginal cells identified by the above-mentioned findings, the effects of neurotransmitters and hormones in regulating intracellular Ca2+ concentration ([Ca2+]i) were investigated. The addition of 10(-4) M ATP caused an increase in [Ca2+]i, which was independent of the presence of extracellular Ca2+. The rank order of agonist potency was ATP > alpha, beta-methylene ATP > ADP. Adenosine, however, evoked no response. Cultured marginal cells may possess P2-purinergic receptors which mobilize Ca2+ from the intracellular Ca2+ pool.
通过培养细胞的形态学、免疫组织化学和生理学特征,建立并确认了豚鼠血管纹外植体边缘细胞的原代培养。细胞角蛋白18呈阳性染色,波形蛋白和结蛋白无表达,表明培养的多边形细胞起源于上皮。培养细胞的电子显微镜观察结果类似于边缘细胞的形态特征。向培养细胞中添加100微摩尔哇巴因可显著提高细胞内钠离子浓度,为钠泵的存在提供了证据。利用上述结果鉴定的培养边缘细胞,研究了神经递质和激素对调节细胞内钙离子浓度([Ca2+]i)的影响。添加10^(-4) M ATP可导致[Ca2+]i升高,这与细胞外钙离子的存在无关。激动剂效力的顺序为ATP>α,β-亚甲基ATP>ADP。然而,腺苷未引起反应。培养的边缘细胞可能具有P2-嘌呤能受体,可从细胞内钙库中动员钙离子。