Full and Partial Agonists of Muscarinic M3 Receptors Reveal Single and Oscillatory Ca2+ Responses by β2-Adrenoceptors

Abstract

Under physiological circumstances, cellular responses often reflect integration of signaling by two or more different receptors activated coincidentally or sequentially. In addition to heterologous desensitization, there are examples in which receptor activation either reveals or potentiates signaling by a different receptor type, although this is perhaps less well explored. Here, we characterize one such interaction between endogenous receptors in human embryonic kidney 293 cells in which Galpha(q/11)-coupled muscarinic M(3) receptors facilitate Ca(2+) signaling by Galpha(s)-coupled beta(2)-adrenoceptors. Measurement of changes in intracellular [Ca(2+)] demonstrated that noradrenaline released Ca(2+) from thapsigargin-sensitive intracellular stores only during activation of muscarinic receptors. Agonists with low efficacy for muscarinic receptor-mediated Ca(2+) responses facilitated cross-talk more effectively than full agonists. The cross-talk required Galpha(s) and was dependent upon intracellular Ca(2+) release channels, particularly inositol (1,4,5)-trisphosphate receptors. However, beta(2)-adrenoceptor-mediated Ca(2+) release was independent of measurable increases in phospholipase C activity and resistant to inhibitors of protein kinases A and C. Interestingly, single-cell imaging demonstrated that particularly lower concentrations of muscarinic receptor agonists facilitated marked oscillatory Ca(2+) signaling to noradrenaline. Thus, activation of muscarinic M(3) receptors profoundly influences the magnitude and oscillatory behavior of intracellular Ca(2+) signaling by beta(2)-adrenoceptors. Although these receptor subtypes are often coexpressed and mediate contrasting acute physiological effects, altered oscillatory Ca(2+) signaling suggests that cross-talk could influence longer term events through, for example, regulating gene transcription.