ACELL Apr. 45/4

Seo, J. T., H. Sugiya, S. I. Lee, M. C. Steward, and A. C. Elliott. Caffeine does not inhibit substance P-evoked intracellular Ca21 mobilization in rat salivary acinar cells. Am. J. Physiol. 276 (Cell Physiol. 45): C915–C922, 1999.—We used the Ca21-sensitive fluorescent dye fura 2, together with measurements of intracellular D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], to assess the inhibitory effects of caffeine on signal transduction via G protein-coupled receptor pathways in isolated rat mandibular salivary acinar cells. ACh, norepinephrine (NE), and substance P (SP) all evoked substantial increases in the intracellular free Ca21 concentration ([Ca]i). Responses to ACh and NE were markedly inhibited by prior application of 20 mM caffeine. The inhibitory effect of caffeine was not reproduced by phosphodiesterase inhibition with IBMX or addition of cell-permeant dibutyryl cAMP. In contrast to the ACh and NE responses, the [Ca]i response to SP was unaffected by caffeine. Despite this, SP and ACh appeared to mobilize Ca21 from a common intracellular pool. Measurements of agonist-induced changes in Ins(1,4,5)P3 levels confirmed that caffeine inhibited the stimulus-response coupling pathway at a point before Ins(1,4,5)P3 generation. Caffeine did not, however, inhibit [Ca]i responses evoked by direct activation of G proteins with 40 mM F2. These data show that caffeine inhibits G protein-coupled signal transduction in these cells at some element that is common to the muscarinic and a-adrenergic signaling pathways but is not shared by the SP signaling pathway. We suggest that this element might be a specific structural motif on the G protein-coupled muscarinic and a-adrenergic receptors.