Characteristics of a rat cortical collecting duct cell line that maintains high transepithelial resistance.
Characteristics of a rat cortical collecting duct cell line that maintains high transepithelial resistance.
复制标题
维持高跨上皮阻力的大鼠皮质集合管细胞系的特征。
作者：
Marcel Blot;Marcel Blot;Monique Laplace;Monique Laplace;Françoise Cluzeaud;Françoise Cluzeaud;Claudia Capurro;Claudia Capurro;R. Cassingéna;R. Cassingéna;Alain Vandewalle;Alain Vandewalle;Nicolette Farman;Nicolette Farman;J. Bonvalet;J. Bonvalet
This study describes the establishment of a rat kidney cortical collecting duct (CCD) clonal cell line (RCCD1 cells) that maintains high transepithelial resistance and specific hormonal sensitivities. Immortalized cells were obtained by infection of primary cultured CCD cells with the wild-type simian virus 40. Grown on Petri dishes, RCCD1 cells are organized as monolayers of cuboid cells separated by tight junctions and form domes. Grown on permeable filters, confluent RCCD1 cells exhibit high transepithelial resistance (Rt: 2390 +/- 140 omega. cm2), transepithelial potential difference (PD) of -10.5 +/- 1.2 mV lumen negative, an associated short-circuit current (Isc) of 4.3 +/- 0.5 microA/cm2, and generated significant Na+, K+, H+ and HCO3- gradients, reflecting Na+ and H+ reabsorption and K+ and HCO3- secretion. RCCD1 cells exhibit features of both principal (PC) and intercalated (IC) cells. Consistent with PC phenotype, about 50% of the cells were positively stained by a PC-specific agglutinin. In situ hybridization studies revealed the presence of alpha, beta and gamma subunit mRNAs of the amiloride-sensitive epithelial Na+ channel and alpha 1 and beta 1 subunits of Na(+)-K(+)-ATPase. Moreover, Na(+)-K(+)-ATPase was immunolocalized at the basolateral side of the cells. Arginine vasopressin (AVP) induced a significant increase in both cellular cAMP content and Isc. Amiloride decreased in a dose-dependent manner Isc from untreated and AVP-treated RCCD1 cells. In addition, a barium-sensitive K+ conductance was evidenced in the apical side of the cells. Consistent with IC phenotype, isoproterenol (ISO) provoked a large increase in cellular cAMP and stimulated Isc. The effect of ISO on Isc was blocked by 5 x 10(-3) M DPC, a chloride channel blocker. Finally, AVP plus ISO had additive effect on Isc. Taken together, these results provide evidence that the RCCD1 cell line has maintained many of the original properties of rat CCD from which they were derived.
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大鼠肾集合管中的三种不同的细胞群。
DOI：
10.1152/ajpcell.1987.253.2.c323
发表时间：
1987
期刊：
The American journal of physiology
影响因子：
--
作者：
Holthöfer,H;Schulte,BA;Pasternack,G;Siegel,GJ;Spicer,SS
通讯作者：
Spicer,SS
影响因子：
19.6
作者：
Stokes,JB
通讯作者：
Stokes,JB
DOI：
10.1152/ajprenal.1990.258.2.f273
发表时间：
1990
期刊：
The American journal of physiology
影响因子：
--
作者：
Light,DB;Schwiebert,EM;Fejes-Toth,G;Naray-Fejes-Toth,A;Karlson,KH;McCann,FV;Stanton,BA
通讯作者：
Stanton,BA
影响因子：
19.6
作者：
B. Stanton;D. Biemesderfer;J. Wade;G. Giebisch
通讯作者：
B. Stanton;D. Biemesderfer;J. Wade;G. Giebisch
DOI：
10.1016/s0021-9258(18)38060-8
发表时间：
1988-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
D. Felsenfeld;K. Sweadner
通讯作者：
D. Felsenfeld;K. Sweadner