Cole J A, Forte L R, Krause W J, Thorne P K
Department of Pharmacology, School of Medicine, University of Missouri, Columbia.
Am J Physiol. 1989 Apr;256(4 Pt 2):F672-9. doi: 10.1152/ajprenal.1989.256.4.F672.
Three clonal subpopulations of opossum kidney (OK) cells were derived from the parental line. The distribution of apical microvilli suggested that the OK cell line was heterogeneous. The clonal OK sublines appeared homogeneous as reflected by microvilli, which were uniformly distributed on the apical surface. Parathyroid hormone (PTH), forskolin (FSK), and prostaglandin E1 (PGE1) increased adenosine 3',5'-cyclic monophosphate (cAMP) formation in OK cells and all of the clones. PTH inhibited sodium-dependent phosphate transport in parental cells and in OK/B and OK/P clones with maximal effects appearing at 4, 2, and 1 h, respectively. PTH had no effect on phosphate transport in OK/H cells. FSK inhibited phosphate transport in parental cells and OK/B and OK/P clones but was relatively ineffective in OK/H cells. PGE1 decreased phosphate transport in OK/B and OK/P cells but was ineffective in the parental line and in OK/H cells. Phorbol 12-myristate 13-acetate, a potent inhibitor of phosphate transport in the parental OK cell line, had little effect in the clonal sublines. These clonal lines have remained phenotypically stable for 10 passages and should prove useful in studying the regulation of phosphate transport by PTH as well as addressing the question of whether PTH receptor subclasses exist which couple to cAMP and/or calcium effector systems in kidney cells.
从亲本细胞系中衍生出了负鼠肾(OK)细胞的三个克隆亚群。顶端微绒毛的分布表明OK细胞系是异质性的。克隆的OK亚系在微绒毛方面表现出均一性,微绒毛均匀分布在顶端表面。甲状旁腺激素(PTH)、福斯可林(FSK)和前列腺素E1(PGE1)可增加OK细胞及其所有克隆中的3',5'-环磷酸腺苷(cAMP)生成。PTH抑制亲本细胞以及OK/B和OK/P克隆中钠依赖性磷酸盐转运,最大效应分别在4小时、2小时和1小时出现。PTH对OK/H细胞中的磷酸盐转运没有影响。FSK抑制亲本细胞以及OK/B和OK/P克隆中的磷酸盐转运,但在OK/H细胞中相对无效。PGE1降低OK/B和OK/P细胞中的磷酸盐转运,但在亲本细胞系和OK/H细胞中无效。佛波酯12-肉豆蔻酸酯13-乙酸酯是亲本OK细胞系中磷酸盐转运的有效抑制剂,在克隆亚系中作用很小。这些克隆系在传代10次后表型仍保持稳定,在研究PTH对磷酸盐转运的调节以及解决肾细胞中是否存在与cAMP和/或钙效应系统偶联的PTH受体亚类问题方面应会很有用。