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Titolo:
CHARACTERISTICS OF A RAT CORTICAL COLLECTING DUCT CELL-LINE THAT MAINTAINS HIGH TRANSEPITHELIAL RESISTANCE
Autore:
BLOTCHABAUD M; LAPLACE M; CLUZEAUD F; CAPURRO C; CASSINGENA R; VANDEWALLE A; FARMAN N; BONVALET JP;
Indirizzi:
UNIV PARIS 07,INSERM,INST FEDERAT RECH CELLULES EPITHELIALES,U246,BP 416 F-75870 PARIS 18 FRANCE GENET MOLEC LAB,CNRS UPR 42 VILLEJUIF FRANCE
Titolo Testata:
Kidney international
fascicolo: 2, volume: 50, anno: 1996,
pagine: 367 - 376
SICI:
0085-2538(1996)50:2<367:COARCC>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
NA+-K+-ATPASE; INSITU HYBRIDIZATION; EPITHELIAL-CELLS; PRIMARY CULTURES; SIMIAN VIRUS-40; PRINCIPAL CELLS; RABBIT KIDNEY; TRANSPORT; TUBULE; EXPRESSION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
41
Recensione:
Indirizzi per estratti:
Citazione:
M. Blotchabaud et al., "CHARACTERISTICS OF A RAT CORTICAL COLLECTING DUCT CELL-LINE THAT MAINTAINS HIGH TRANSEPITHELIAL RESISTANCE", Kidney international, 50(2), 1996, pp. 367-376

Abstract

This study describes the establishment of a rat kidney cortical collecting duct (CCD) clonal cell line (RCCD(1) 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, RCCD(1) cells are organized as monolayers of cuboid cells separated by tight junctions and form domes. Grown on permeable filters, confluent RCCD(1) cells exhibit high transepithelial resistance (R(t): 2390 +/- 140 Ohm . cm(2)), transepithelial potential difference (PD) of -10.5 +/- 1.2 mV lumen negative, an associated short-circuit current (Isc) of 4.3 +/- 0.5 mu A/cm(2), and generated significant Na+, K+, H+ and HCO3- gradients, reflecting Na+ and H+ reabsorption and K+ and HCO3- secretion. RCCD(1) 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 RCCD(1) 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 RCCD(1) cell line has maintained many of the original properties of rat CCD from which they were derived.

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Documento generato il 27/11/20 alle ore 22:15:52