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Am J Physiol Cell Physiol 280: C1193-C1203, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 5, C1193-C1203, May 2001

Nifedipine-activated Ca2+ permeability in newborn rat cortical collecting duct cells in primary culture

Laura Valencia1, Michel Bidet2, Sonia Martial2, Elsa Sanchez1, Estela Melendez1, Michel Tauc2, Chantal Poujeol2, Dolores Martin1, Maria Del Carmen Namorado1, Jose Luis Reyes1, and Philippe Poujeol2

1 Departamento de Fisiología, Centro de Investigación y de Estudios Avanzados del Institúto Politécnico Nacional, Mexico City DF 07000, Mexico; and 2 Unite Mixte de Recherche-Centre National de la Recherche Scientifique 6548, Université de Nice-Sophia Antipolis, O6108 Nice Cedex 2, France

To characterize Ca2+ transport in newborn rat cortical collecting duct (CCD) cells, we used nifedipine, which in adult rat distal tubules inhibits the intracellular Ca2+ concentration ([Ca2+]i) increase in response to hormonal activation. We found that the dihydropyridine (DHP) nifedipine (20 µM) produced an increase in [Ca2+]i from 87.6 ± 3.3 nM to 389.9 ± 29.0 nM in 65% of the cells. Similar effects of other DHP (BAY K 8644, isradipine) were also observed. Conversely, DHPs did not induce any increase in [Ca2+]i in cells obtained from proximal convoluted tubule. In CCD cells, neither verapamil nor diltiazem induced any rise in [Ca2+]i. Experiments in the presence of EGTA showed that external Ca2+ was required for the nifedipine effect, while lanthanum (20 µM), gadolinium (100 µM), and diltiazem (20 µM) inhibited the effect. Experiments done in the presence of valinomycin resulted in the same nifedipine effect, showing that K+ channels were not involved in the nifedipine-induced [Ca2+]i rise. H2O2 also triggered [Ca2+]i rise. However, nifedipine-induced [Ca2+]i increase was not affected by protamine. In conclusion, the present results indicate that 1) primary cultures of cells from terminal nephron of newborn rats are a useful tool for investigating Ca2+ transport mechanisms during growth, and 2) newborn rat CCD cells in primary culture exhibit a new apical nifedipine-activated Ca2+ channel of capacitive type (either transient receptor potential or leak channel).

calcium channel; dihydropyridine; kidney; newborn


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