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J Biol Chem, Vol. 273, Issue 8, 4695-4704, February 20, 1998

Coexpression of Ligand-gated P_2X and G Protein-coupled P_2Y Receptors in Smooth Muscle
PREFERENTIAL ACTIVATION OF P_2Y RECEPTORS COUPLED TO PHOSPHOLIPASE C (PLC)-1 VIA G_q/11 AND TO PLC-3 VIA G_i3

From the Departments of Medicine and Physiology, Medical College of Virginia, Richmond, Virginia 23298-0711

P₂ receptor subtypes and their signaling mechanisms were characterized in dispersed smooth muscle cells. UTP and ATP stimulated inositol 1,4,5-triphosphate formation, Ca²⁺ release, and contraction that were abolished by U-73122 and guanosine 5'-O-(3-thio)diphosphate, and partly inhibited (50-60%) by pertussis toxin (PTX). ATP analogs (adenosine 5'-(,-methylene)triphosphate, adenosine 5'-(,-methylene)triphosphate, and 2-methylthio-ATP) stimulated Ca²⁺ influx and contraction that were abolished by nifedipine and in Ca²⁺-free medium. Micromolar concentrations of ATP stimulated both Ca²⁺ influx and Ca²⁺ release.

ATP and UTP activated G_q/11 and G_i3 in gastric and aortic smooth muscle and heart membranes, G_q/11 and G_i1 and/or G_i2 in liver membranes, and G_o and G_i1-3 in brain membranes. Phosphoinositide hydrolysis stimulated by ATP and UTP was mediated concurrently by G_q/11-dependent activation of phospholipase (PL) C-1 and G_i3-dependent activation of PLC-3. Phosphoinositide hydrolysis was partially inhibited by PTX or by antibodies to G_q/11, G, PLC-1, or PLC-3, and completely inhibited by the following combinations (PLC-1 and PLC-3 antibodies; G_q/11 and G antibodies; PLC-1 and G antibodies; PTX with either PLC-1 or G_q/11 antibody).

The pattern of responses implied that P_2Y2 receptors in visceral, and probably vascular, smooth muscle are coupled to PLC-1 via G_q/11 and to PLC-3 via G_i3. These receptors co-exist with ligand-gated P_2X1 receptors activated by ATP analogs and high levels of ATP.

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