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Vol. 60, Issue 3, 514-520, September 2001

cAMP Modulates Exocytotic Kinetics and Increases Quantal Size in Chromaffin Cells

José D. Machado, Araceli Morales, José F. Gomez, and Ricardo Borges

Unidad de Farmacología (J.D.M., J.F.G., R.B.) and Laboratorio de Neurobiología Celular (A.M), Facultad de Medicina, Universidad de La Laguna, Tenerife, Spain

The role of cAMP/cAMP-dependent protein kinase (PKA) on the late phase of exocytosis has been studied by amperometry on Ba2+-stimulated single bovine chromaffin cells. Forskolin (FSK) increases the intracellular cAMP levels in a concentration-dependent manner. Forskolin (100 nM) does not increase the number of exocytotic events, although it significantly increases the net granule content of catecholamines (CA), which is accompanied by a slowing of the process of degranulation. These effects are reversible, occur within 15 to 60 s, and are not due to newly synthesized CA. Isoprenaline, pituitary adenylate cyclase-activating polypeptide-38 or dB-cAMP reproduce FSK effects as does cholera toxin. The inhibition of phosphodiesterases with 3-isobutyl-1-methylxanthine mimics and potentiates the effect of FSK and isoprenaline. Rolipram and okadaic acid also produce a drastic increase in net granule content of CA, whereas H-89 attenuates the FSK response. These data indicate that cyclic AMP/PKA might favor the granule aggregation before its fusion with cell membrane and slow the late step of the exocytotic process.

Copyright © 2001 by The American Society for Pharmacology and Experimental Therapeutics


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