Vasopressin-stimulated chloride transport in transimmortalized mouse cell lines derived from the distal convoluted tubule and cortical and inner medullary collecting ducts.

摘要

BACKGROUND: The fine control of NaCl absorption takes place in the distal parts of the renal tubule, but the regulation of Cl(-) transport in this region has not been fully elucidated. We have analysed the effects of dD-arginine vasopressin (dDAVP) on Cl(-) fluxes in cultured mouse distal convoluted tubule (mpkDCT), cortical collecting duct (mpkCCD) and inner medullary collecting duct (mpkIMCD) cell lines. METHODS: RT-PCR and Western blotting were used to detect the amiloride-sensitive sodium channel (ENaC) and cystic fibrosis transmembrane conductance regulator (CFTR) mRNAs and protein in cultured mpkDCT, mpkCCD and mpkIMCD cells. Cl(-) fluxes were analysed by measuring the short-circuit current (I(sc)) and bidirectional (36)Cl(-) fluxes on confluent cells grown on filters. RESULTS: All three cell lines expressed ENaC and CFTR and had I(sc) stimulated by dDAVP. The rise in I(sc) caused by dDAVP (10(-8) M) was inhibited by amiloride, and to a lesser extent by 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) in all three cell lines. The dDAVP-dependent I(sc) measured under apical Na(+)-free condition was reduced by Cl(-) channel blockers with a profile (NPPB>glibenclamide>DIDS), similar to that for rat CFTR. dDAVP stimulated the apical-to-basal (36)Cl(-) flux and to a lesser extent the basal-to-apical (36)Cl(-) flux under open-circuit condition in all three cultured cell lines. Adding NPPB to the apical side reduced the basal-to-apical (36)Cl(-) flux but not the opposite (36)Cl(-) flux from dDAVP-treated cells. CONCLUSION: These results indicate that dDAVP stimulates the bi-directional flux of Cl(-), resulting in net Cl(-)absorption, in these cultured mouse distal and collecting duct cells. I(sc) experiments also suggest the presence of a minor component of electrogenic Cl(-) secretion, possibly mediated by CFTR.