Abstract
The effects of G-protein activation were investigated on tonic, large depolarization-induced Ca2+ channel facilitation in cultured bovine adrenal chromaffin cells. Under whole-cell voltage clamp, activation of G proteins by intracellular dialysis with 200 μM GTP-γS did not significantly affect prepulse facilitation or whole-cell Ba2+ current (I Ba) density. In contrast, inactivation of G proteins by intracellular GDP-βS or pertussis toxin (PTX) pretreatment completely abolished or markedly attenuated facilitation of I Ba, respectively. GDP-βS dialysis resulted in nearly a threefold increase in peak I Ba density, whereas PTX pretreatment resulted in a 50% increase. Our results indicate that under control recording conditions (200 μm intracellular GTP), G proteins are tonically activated and suppress high-voltage-activated (HVA) Ca2+ channels in a voltage-dependent and voltage-independent manner. Local superfusion of chromaffin cells with normal bath solution produced a rapid and reversible increase (∼50%) in I Ba amplitudes that also abolished prepulse facilitation. Together, these results demonstrate that tonic facilitation of HVA Ca2+ channels in bovine chromaffin cells involves the voltage-dependent relief of a G-protein-mediated suppression, imposed by chromaffin cell secretory products that feedback and activate G-protein-coupled autoreceptors.
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This work was supported by a National Science Foundation grant (DCB-8812562), American Heart Association-Ohio Affiliate grant (SW-91-18), and an American Parkinson's Disease Association grant. C.A.D. was supported by a predoctoral National Research Service Award (National Institutes of Health training grant HL07571-08). The authors thank Kluener's Packing Co. for their generous supply of adrenal glands.
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Doupnik, C.A., Pun, R.Y.K. G-Protein activation mediates prepulse facilitation of Ca2+ channel currents in bovine chromaffin cells. J. Membarin Biol. 140, 47–56 (1994). https://doi.org/10.1007/BF00234485
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DOI: https://doi.org/10.1007/BF00234485