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- Title
Characteristics of a rat cortical collecting duct cell line that maintains high transepithelial resistance.
- Authors
Blot-Chabaud, Marcel; Laplace, Monique; Cluzeaud, Françoise; Capurro, Claudia; Cassingeña, Roland; Vandewalle, Alain; Farman, Nicolette; Bonvalet, Jean Pierre
- Abstract
Characteristics of a rat cortical collecting duct cell line that maintains high transepithelial resistance. 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 RCCD I cells exhibit high transepithelial resistance (R1: 239U &plusm; 140 ω · em2), transepithelial potential difference (PD) of -10.5 ± 1.2 mV lumen negative, an associated short-circuit current (Isc) of 4.3 ± 0.5μA/cm2-. And generated significant Na +, K +, H + and HCO3, gradients, reflecting Na+ and H+ reabsorption and K+ and HCO3-, secretion. RCCD, 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 α1,β1 subunit mRNAs of the amiloride-sensitive epithelial Na+ channel and α+; and subunits of N+-K+-ATPasc. Moreover, Na+-K+-ATPasc 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 RCC'D, cells. In addition, a bariumsensitive K+ conductance was evidenced in the apical side of the cells. Consistent with IC phenotype. Isoproterenol (ISO) provoked a large increase in cellular cAMPand stimulated Isc. The effect of ISC) on Isc was blocked by 5 × H-3) M DPC, a chloride channel blocker. Finally. AVP plus ISO bad 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.
- Subjects
KIDNEY cortex; CELL lines; CELL culture; LABORATORY rats; KIDNEYS; NEPHROLOGY; INTERNAL medicine
- Publication
Kidney International, 1996, Vol 50, Issue 2, p367
- ISSN
0085-2538
- Publication type
Article
- DOI
10.1038/ki.1996.325