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Endocrinology Vol. 138, No. 8 3299-3307
Copyright © 1997 by The Endocrine Society

ARTICLES

Association of the G Protein {alpha}q/{alpha}11-Subunit with Cytoskeleton in Adrenal Glomerulosa Cells: Role in Receptor-Effector Coupling1

Mylène Côté, Marcel D. Payet, Marie-Noëlle Dufour, Gilles Guillon and Nicole Gallo-Payet2

Service of Endocrinology, Department of Medicine (M.C., N.G.-P.), and the Department of Physiology and Biophysics (M.D.P.), Faculty of Medicine, University of Sherbrooke, Sherbrooke, Quebec, Canada J1H 5N4; and INSERM U-401 (M.-N.D., G.G.), Montpellier, France

Address all correspondence and requests for reprints to: Dr. Nicole Gallo-Payet, Service of Endocrinology, Department of Medicine, Faculty of Medicine, University of Sherbrooke, 3001 12th Ave North, Sherbrooke, Quebec, Canada J1H 5N4. E-mail:n.gallo{at}courrier.usherb.ca

In 3-day primary cultures of rat glomerulosa cells, a 30-min preincubation with either 10 µM colchicine (a microtubule-disrupting agent) or 10 µM cytochalasin B (a microfilament-disrupting agent) decreased angiotensin II (Ang II)-induced inositol phosphate accumulation by 50%. Moreover, both drugs decreased inositol phosphate production induced by fluoroaluminate (a nonspecific activator of all G proteins), indicating that both microtubules and microfilaments are essential for phospholipase C activation. Analysis of microfilament- and microtubule-enriched fractions and immunoprecipitation of actin and tubulin revealed that the {alpha}q/{alpha}11-subunit of the Gq/11 protein was associated with both structures. Ang II stimulation induced a rapid translocation of {alpha}q/{alpha}11, microfilaments, and microtubules to the membrane and induced a time-dependent increase in the level of {alpha}q/{alpha}11 associated with both microfilaments and microtubules. Moreover, double immunofluorescence staining clearly showed a colocalization of the {alpha}q/{alpha}11-subunit of the Gq/11 coupling protein and microfilament distribution. These associations and plasma membrane redistribution under Ang II stimulation indicate that microfilaments and microtubules are both involved in phospholipase C activation and inositol phosphate production. Moreover, our results indicate that the {alpha}q/{alpha}11 protein is closely associated with cytoskeletal elements and is found both at the plasma membrane level as well as on intracellular stress fibers.




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