PHENOTYPICALLY AND KARYOTYPICALLY DISTINCT MADIN-DARBY CANINE KIDNEY-CELL CLONES RESPOND DIFFERENTLY TO ALKALINE STRESS

被引:54
作者
WUNSCH, S [1 ]
GEKLE, M [1 ]
KERSTING, U [1 ]
SCHURICHT, B [1 ]
OBERLEITHNER, H [1 ]
机构
[1] UNIV WURZBURG,DEPT PHYSIOL,D-97070 WURZBURG,GERMANY
关键词
D O I
10.1002/jcp.1041640121
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We isolated two cell clones from the wild-type Madin-Darby canine kidney cell line (MDCK) that resembles renal collecting duct epithelium. Morphology and karyotypes of the two cell clones were evaluated. The MDCK-C7 cell clone morphologically resembles principal cells (polygonal cell shape, flat), while the MDCK-C11 clone resembles intercalated cells (cuboidal cell shape, high). The diploid chromosome number of MDCK-C7 cells is 83.1 +/- 0.2 (n = 139); that for MDCK-C11 cells is 78.8 +/- 0.1 (n = 128). Culture of MDCK-C7 cells in alkaline medium (pH 7.7) induced irreversible phenotypical and genotypical alterations. Transformed MDCK-C7F cells are characterized by two abnormal (biarmed) chromosomes. In contrast, MDCK-C11 cells are not phenotypically altered by alkaline stress. In order to elucidate the role of intracellular pH (pH(i)) in the transformation process, we measured pH(i) under control conditions (pH 7.4), after 5 min exposure to alkaline stress (''acute experiment,'' pH 7.7) and after incubation of the cells in alkaline medium for two weeks (''chronic experiment,'' pH 7.7). Under control conditions, MDCK-C7 cells maintained pH(i) at 7.14 +/- 0.01 (n = 154) and MDCK-C11 cells at 7.01 +/- 0.01 (n = 147). Acute alkaline stress increased pH(i) of both cell types to similar steady-state values. Under chronic alkaline stress, MDCK-C7 cells were unable to maintain intracellular pH within normal limits exhibiting sustained alkalinization, whereas MDCK-C11 cells could successfully regulate pH(i). We conclude that wild-type MDCK cells consist of two genetically distinct subpopulations with different morphology and function. Only the MDCK-C7 clone that resembles the principle cell type of renal collecting duct can be transformed by alkaline stress while the MDCK-C11 clone resists this treatment, due to efficient pH(i) control mechanisms. (C) 1995 Wiley-Liss, Inc.
引用
收藏
页码:164 / 171
页数:8
相关论文
共 30 条
  • [1] Gaush R.C., Walter W.L., Smith F., Characterization of an established line of canine kidney cells (MDCK), Proc. Soc. Exp. Biol. Med., 122, pp. 931-935, (1966)
  • [2] Gekle M., Wunsch S., Oberleithner H., Silbernagl S., Characterization of two MDCK‐cell INFtypes as a model system to study principal and intercalated cell properties, Pflügers Arch., 428, pp. 157-162, (1994)
  • [3] Gillies R.J., Martinez-Zeguilan R., Peterson E.P., Perona R, Role of intracellular pH in mammalian cell proliferation, Cellular Physiology and Biochemistry, 2, pp. 159-179, (1992)
  • [4] Grynkiewicz G., Poenie M., Tsien R.Y., A new generation of Ca<sup>2+</sup> indicators with greatly improved fluorescent properties, J. Biol. Chem., 260, pp. 3440-3450, (1985)
  • [5] Hesketh T.R., Moore J.P., Morris J.D.H., Taylor M.V., Rogers J., Smith G.A., Metcalf J.C., A common sequence of calcium and pH signals in the mitogenic stimulation of eukaryotic cells, Nature, 313, pp. 481-484, (1985)
  • [6] Kersting U., Joha H., Steigner W., Gabner B., Gstraunthaler G., Pfaller W., Oberleithner H., Fusion of cultured dog kidney (MDCK) cells. I. Technique, fate of plasma membrane and cell nuclei, J. Membr, Biol., 111, pp. 37-48, (1988)
  • [7] Kersting U., Schwab A., Treidtel M., Pfaller W., Gstraunthaler G., Steigner W., Oberleithner H., Differentiation of Madin‐Darby canine kidney cells depends on cell culture conditions, Cell. Physiol. Biochem., 3, pp. 42-55, (1993)
  • [8] Kurtz I., Golchini K., Na<sup>+</sup>‐independent Cl<sup>−</sup>/HCO<sub>3</sub><sup>−</sup> exchange in Madin‐Darby canine kidney cells, J. Biol. Chem., 262, pp. 4516-4521, (1987)
  • [9] Manolache M., Ross W.M., Schmid M., Banding analysis of the somatic chromosomes of the domestic dog (canis familiaris), Can. J. Genet. Cytol., 18, pp. 513-518, (1976)
  • [10] Meier K.E., Snavely M.D., Brown S.L., Brown J.H., Insel P.A., α <sub>1</sub>‐ and b<sub>1</sub>‐adrenergic receptor expression in the Madir‐Darby canine kidney epithelial cell line, J. Cell. Biol., 97, pp. 405-415, (1983)