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Endocrinology Vol. 141, No. 2 684-692
Copyright © 2000 by The Endocrine Society

ARTICLES

A Ras-Dependent Chloride Current Activated by Adrenocorticotropin in Rat Adrenal Zona Glomerulosa Cells1

Alzbeta Chorvátová2, Louis Gendron, Lyne Bilodeau, Nicole Gallo-Payet and Marcel D. Payet

Department of Physiology and Biophysics (A.C., L.G., L.B., N.G.-P., M.D.P.) and Service of Endocrinology (L.G., N.G.-P.), Faculty of Medicine, Sherbrooke, Québec, J1H 5N4, Canada

Address all correspondence and requests for reprints to: Dr. Marcel Daniel Payet, Department of Physiology and Biophysics, Faculty of Medicine, University of Sherbrooke, 3001, 12th Avenue North, Sherbrooke, Québec, Canada, J1H 5N4. E-mail: mpayet01{at}courrier.usherb.ca

In the present study, we report that ACTH induces a transient chloride current. The lack of correlation between ACTH-induced cAMP production and amplitude of the Cl- current, as well as the absence of stimulation by forskolin or 8Br-cAMP indicated that the ACTH-induced current was not cAMP-dependent. We explored the possibility that one or several elements of the Ras/Raf MAPK cascade were involved. Indeed, we found that ACTH at 10-10 M induced activation of Ras. Inhibition of the current by QEHA peptide, a Gß{gamma} sequestrant, demonstrated that Gß{gamma} subunits transduced the message. Blockage of the Ras activation using an inhibitor of farnesyl transferase (BZA-5B) or the monoclonal antibody H-Ras(259) abrogated the current. Moreover, the addition of Ras-GTP{gamma}S in the pipette medium gave rise to the Cl- current. Treatment of the cells with BZA decreased the aldosterone secretion induced by 10-10 M ACTH but not that induced by 10-8 M ACTH, confirming the involvement of Ras in steroid secretion. We conclude that ACTH triggers a Cl- current through the activation of the Ras protein by Gß{gamma} subunits. This current, activated at physiological ACTH concentrations (1 to 100 pM) where cAMP production is very low, could play a significant role in aldosterone production.




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