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J Biol Chem, Vol. 274, Issue 28, 19943-19948, July 9, 1999

Angiotensin II Negatively Modulates L-type Calcium Channels through a Pertussis Toxin-sensitive G Protein in Adrenal Glomerulosa Cells

Andrés D. Maturana^§, Andrés J. Casal, Nicolas Demaurex^¶, Michel B. Vallotton, Alessandro M. Capponi, and Michel F. Rossier

From the  Division of Endocrinology and Diabetology, Department of Internal Medicine, the  Laboratory of Clinical Chemistry, Department of Pathology, the ^¶ Department of Physiology, and the ^§ Fondation pour Recherches Médicales, University Hospital, 24 rue Micheli-du-Crest,CH-1211 Geneva 14, Switzerland

In bovine adrenal glomerulosa cells, angiotensin II and extracellular K⁺ stimulate aldosterone secretion in a calcium-dependent manner. In these cells, physiological concentrations of extracellular potassium activate both T-type (low threshold) and L-type (high threshold) voltage-operated calcium channels. Paradoxically, the cytosolic calcium response to 9 mM K⁺ is inhibited by angiotensin II. Because K⁺-induced calcium changes observed in the cytosol are almost exclusively due to L-type channel activity, we therefore studied the mechanisms of L-type channel regulation by angiotensin II. Using the patch-clamp method in its perforated patch configuration, we observed a marked inhibition (by 63%) of L-type barium currents in response to angiotensin II. This effect of the hormone was completely prevented by losartan, a specific antagonist of the AT₁ receptor subtype. Moreover, this inhibition was strongly reduced when the cells were previously treated for 1 night with pertussis toxin. An effect of pertussis toxin was also observed on the modulation by angiotensin II of the K⁺ (9 mM)-induced cytosolic calcium response in fura-2-loaded cells, as well as on the angiotensin II-induced aldosterone secretion, at both low (3 mM) and high (9 mM) K⁺ concentrations. Finally, the expression of both G_o and G_i proteins in bovine glomerulosa cells was detected by immunoblotting. Altogether, these results strongly suggest that in bovine glomerulosa cells, a pertussis toxin-sensitive G protein is involved in the inhibition of L-type channel activity induced by angiotensin II.

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