In order to probe the nature of oxytocin receptor (OTR)/G alpha(q/11) protein coupling, we examined the effect of co-expression of OTR intracellular domains on oxytocin-stimulated phosphoinositide turnover in COSM6 cells overexpressing OTR and G alpha(q). Co-expression of G alpha(q) enhanced the oxytocin response maximally at a pOTR/pG alpha(q) plasmid transfection ratio of 1:0.16. In cells co-expressing OTR and G alpha(q/11), oxytocin stimulated phosphoinositide turnover with an EC50 of 48 nM. Co-transfection with plasmids expressing OTR intracellular domains inhibited oxytocin-stimulated phosphoinositide turnover by 23 +/- 6% (1i), 37 +/- 4% (2i), 55 +/- 6% (3i), and 40 +/- 6% (4i), respectively (P < 0.01). Expression of the 3i loop of the alpha(1B)-adrenergic receptor, which also couples to G alpha(q/11), inhibited phosphoinositide turnover by 35 +/- 2% (P < 0.01), while expression of the 3i loop of the dopamine 1A receptor, which couples to G alpha(s), had no effect. While these data indicate a functional role for the OTR 3i loop, they also suggest that interactions with more than one intracellular domain probably mediate the coupling of OTR to the G alpha(q/11) class of GTP-binding proteins.
Amino Acid Sequence Animals