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J Physiol Volume 506, Number 2, 319-329, January 15, 1998
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The Journal of Physiology (1998), 506.2, pp. 319-329
© Copyright 1998 The Physiological Society

On the role of endogenous G-protein betagamma subunits in N-type Ca2+ current inhibition by neurotransmitters in rat sympathetic neurones

Patrick Delmas, David A. Brown, Mariza Dayrell, Fe C. Abogadie, Malcolm P. Caulfield * and Noel J. Buckley

Wellcome Laboratory for Molecular Pharmacology , Department of Pharmacology, University College London, Gower Street, London WC1E 6BT and * Neurosciences Institute, Department of Pharmacology and Clinical Pharmacology, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, UK

  1. Using whole-cell and perforated-patch recordings, we have examined the part played by endogenous G-protein betagamma subunits in neurotransmitter-mediated inhibition of N-type Ca2+ channel current (ICa) in dissociated rat superior cervical sympathetic neurones.

  2. Expression of the C-terminus domain of beta-adrenergic receptor kinase 1 (betaARK1), which contains the consensus motif (QXXER) for binding Gbetagamma, reduced the fast (pertussis toxin (PTX)-sensitive) and voltage-dependent inhibition of ICa by noradrenaline and somatostatin, but not the slow (PTX-insensitive) and voltage-independent inhibition induced by angiotensin II. betaARK1 peptide reduced GTP-gamma-S-induced voltage-dependent and PTX-sensitive inhibition of ICa but not GTP-gamma-S-mediated voltage-independent inhibition.

  3. Overexpression of Gbeta1gamma2, which mimicked the voltage-dependent inhibition by reducing ICa density and enhancing basal facilitation, occluded the voltage-dependent noradrenaline- and somatostatin-mediated inhibitions but not the inhibition mediated by angiotensin II.

  4. Co-expression of the C-terminus of betaARK1 with beta1 and gamma2 subunits prevented the effects of Gbetagamma dimers on basal Ca2+ channel behaviour in a manner consistent with the sequestering of Gbetagamma.

  5. The expression of the C-terminus of betaARK1 slowed down reinhibition kinetics of ICa following conditioning depolarizations and induced long-lasting facilitation by cumulatively sequestering betagamma subunits.

  6. Our findings identify endogenous Gbetagamma as the mediator of the voltage-dependent, PTX-sensitive inhibition of ICa induced by both noradrenaline and somatostatin but not the voltage-independent, PTX-insensitive inhibition by angiotensin II. They also support the view that voltage-dependent inhibition results from a direct Gbetagamma-Ca2+ channel interaction.



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