Endothelial G Protein β-Subunits Trigger Nitric Oxide– but not Endothelium-Derived Hyperpolarizing Factor–Dependent Dilation in Rabbit Resistance Arteries

A single subtype of heterotrimeric G protein-coupled receptor controls both nitric oxide (NO) (sensitive to L-arginine analogues) and endothelium-derived hyperpolarizing factor (EDHF) (sensitive to high-external K(+) and apamine) production by the vascular endothelium leading to dilation. We hypothesized that alpha- and betagamma-subunits of the G protein serve as distinct intermediates to produce NO and EDHF. In pressurized resistance arteries, selective pinocytotic endothelial incorporation of specific antibodies (Abs) directed against alpha(q/11)-subunits abolished acetylcholine (Ach)-mediated dilation but failed to influence oxymetazoline (Oxy, alpha(2)-adrenergic receptor agonist)-induced dilation. In contrast, alpha(i1-2)-subunit Abs prevented Oxy- but not Ach-induced dilation. Thus, as expected, endothelial muscarinic and alpha(2)-adrenoceptors couple to G(q) protein and G(i) proteins, respectively. beta-subunit Abs reduced both Ach- and Oxy-induced dilation. The beta-subunit Abs abolished the nitro-L-arginine (L-NNA)-sensitive component but did not impair the high-external K(+)-sensitive component of the dilation induced by Ach and Oxy. Thus, G protein beta-subunits primarily accounted for NO production. Neutralization of Hsp90 and inhibition of the phospholipase C by U73122 (1 micromol/L) or intracellular Ca(2+) buffering with BAPTA-AM (10 micromol/L) sharply reduced NO-dependent but not K(+)-sensitive dilation. In conclusion, mobilization of the G protein beta-subunit is pivotal to NO-dependent dilation triggered through muscarinic and alpha(2)-adrenergic receptors. In contrast, receptor-operated EDHF-dependent dilation was insensitive to beta-subunit Abs. Although not directly activating the NO pathway, alpha-subunit activation is an absolute prerequisite for receptor-operated endothelium-dependent dilation of resistance arteries.