McDermott G F, Hurst R D, Whiteside C I
Department of Medicine, University of Toronto, Ontario, Canada.
Cell Calcium. 1993 May;14(5):387-96. doi: 10.1016/0143-4160(93)90043-6.
The presence of L-type calcium (Ca2+)-channels and the effects of Ca(2+)-channel antagonists on cells of rat glomeruli were investigated. Glomeruli were isolated by graded sieving and after preincubation (10 min) in zero Ca2+, the uptake of 45Ca2+ by glomerular cells was measured. Depolarization with KCl (50 mM) or the dihydropyridine agonist Bay K 8644 (10 microM) stimulated 45Ca2+ uptake by 13% and 24%, respectively, above control (100%), which was inhibited by nifedipine (Nif, 10 microM), P < 0.05, and by both S and R isomers of verapamil (Ver, 10 microM), P < 0.001. In a separate experimental preparation, isolated glomeruli were preloaded (45 min) with 45Ca2+. Following a 45 min perifusion (37 degrees C, CaCl2 1.26 mM, in the absence of 45Ca2+), both KCl (50 mM) and Bay K 8644 (10 microM) induced cellular 45Ca2+ efflux with peak values above control of 11% and 15%, respectively, (P < 0.05). Exposure to Bay K 8644 preceded by depolarization with KCl resulted in enhanced 45Ca2+ efflux identifying the presence of voltage-dependent Ca(2+)-channel activity. Cultured rat mesangial cells grown to confluence on coverslips were preloaded with Fura-2 and cytosolic Ca2+ was measured by microfluorometry. KCl (50 mM), gramicidin (2 microM) and/or Bay K 8644 (6 microM) stimulated Ca2+ influx which was inhibited by Ver (10 microM). Ver did not alter endothelin-stimulated Ca2+ signalling. We conclude that L-type Ca2+ channels are present on both rat glomerular (endothelial and/or mesangial) cells in vivo and on cultured mesangial cells, and their activation may be hormone specific.
