Muscarinic receptor activation of arachidonate-mediated Ca2+ entry in HEK293 cells is independent of phospholipase C

Receptor-enhanced entry of Ca2+ in non-excitable cells is generally ascribed to a capacitative mechanism in which the activation of the entry pathway is specifically dependent on the emptying of agonist-sensitive intracellular Ca2+ stores. Although such entry can be clearly demonstrated under conditions of maximal or near-maximal stimulation, it is uncertain whether such a mechanism can operate during the oscillatory [Ca2+](i) signals that are frequently seen following stimulation with low concentrations of agonists. In this study, we report that the stimulation of human m3 muscarinic receptors stably transfected into HEK293 cells results in the appearance of a novel arachidonate-mediated Ca2+ entry pathway. We show that the generation of arachidonic acid and the activation of this pathway are specifically associated with stimulation at the low agonist concentrations that typically give rise to oscillatory [Ca2+](i) signals. At such agonist concentrations, however, the generation of arachidonic acid is independent of the simultaneous activation of the phospholipase C-inositol 1,4,5-trisphosphate pathway. We further show that the arachidonate-mediated Ca2+ entry demonstrates characteristics that distinguish it from the corresponding capacitative pathway in the same cells and therefore is likely to represent an entirely distinct pathway that is specifically responsible for the receptor-enhanced entry of Ca2+ during [Ca2+], oscillations.