Differential role of Na+/H+ exchange isoforms NHE-1 and NHE-2 in a rat cortical collecting duct cell line.

FORD PAULA; RIVAROLA VALERIA; KIERBEL ARLINET; CHARA OSVALDO; BLOT-CHABAUD MARCEL; FARMAN NICOLETTE; PARISI MARIO; CAPURRO CLAUDIA

JOURNAL OF MEMBRANE BIOLOGY

Springer-Verlag New York inc.

Lugar: New York, USA; Año: 2002 vol. 190 p. 117 - 125

0022-2631

Differential role of Na+/H+ exchange isoforms NHE-1 and NHE-2 in a rat cortical collecting duct cell line. Ford P, Rivarola V, Kierbel A, Chara O, Blot-Chabaud M, Farman N, Parisi M, Capurro C. Laboratorio de Biomembranas, Depto de Fisiologia, Facultad de Medicina, Univ. de Buenos Aires, Paraguay 2155, Piso 7, 1121 Buenos Aires, Argentina. The Na+/H+ exchanger (NHE) constitutes a gene family containing several isoforms that display different membrane localization and are involved in specialized functions. Although basolateral NHE-1 activity was described in the cortical collecting duct (CCD), the localization and function of other NHE isoforms is not yet clear, This study examines the expression, localization, and regulation of NHE isoforms in a rat cortical collecting duct cell line (RCCD1) that has previously been shown to be a good model of CCD cells. Present studies demonstrate the presence of NHE-1 and NHE-2 isoforms, but not NHE-3 and NHE-4, in RCCD1 cells. Cell monolayers, grown on permeable filters, were placed on special holders allowing independent access to apical and basolateral compartments. Intracellular pH (pHi) regulation was spectrofluorometrically studied in basal conditions and after stimulation by NH4Cl acid load or by a hyperosmotic shock. In order to differentiate the roles of NHE-1 and NHE-2, we have used HOE-694, an inhibitor more selective for NHE-1 than for NHE-2. The results obtained strongly suggest that NHE-1 and NHE-2 are expressed in the basolateral membrane but that they have different roles: NHE-1 is responsible for pHi recovery after an acid load and NHE-2 is mainly involved in steady-state pHi and cell volume regulation.