Direct G protein activation reverses impaired CCK signaling in human gallbladders with cholesterol stones.
Human gallbladders were used to investigate the mechanisms of the impaired contraction induced by cholecystokinin (CCK) associated with cholesterol stones. Single muscle cells were isolated enzymatically with collagenase. Inositol 1,4,5-trisphosphate was measured by high-performance liquid chromatography. Diacylglycerol was assayed by thin-layer chromatography. CCK stimulation showed decreased muscle contraction and production of inositol 1,4,5-trisphosphate and diacylglycerol in gallbladders with cholesterol stones compared with those with pigment stones. Exogenous calmodulin induced maximal contraction of 22.4 +/- 0.5 and 21.0 +/- 0.6% in gallbladders with cholesterol and pigment stones, respectively. Similar findings were observed with a synthetic diacylglycerol analogue. Two G protein activators, aluminum fluoride and guanosine 5'-O-(3-thiotriphosphate), evoked similar responses in these two types of gallbladders, with maximal contractions of 21.3 +/- 0.4 and 23.3 +/- 0.5%, respectively, in those with cholesterol stones and 20.9 +/- 0.8 and 22.6 +/- 0.4%, respectively, in those with pigment stones. These results suggest that receptor-dependent ligands like CCK cannot fully activate the intracellular pathways, which, however, can be fully stimulated by circumventing receptors with G protein activators or second messengers. After G protein activation, the pathways appear to be functionally intact. The defect might then reside in the receptor or in the interaction between receptors and G proteins.
D017607:Aluminum Compounds / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002766:Cholecystokinin / Q000502:physiology*; D002769:Cholelithiasis / Q000473:pathology / Q000503:physiopathology*; D002784:Cholesterol / Q000378:metabolism*; D004075:Diglycerides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005459:Fluorides / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000502:physiology*; D005704:Gallbladder / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis; D008297:Male; D008875:Middle Aged; D009119:Muscle Contraction; D015398:Signal Transduction; D012844:Sincalide / Q000494:pharmacology
D017607:Aluminum Compounds; D002147:Calmodulin; D004075:Diglycerides; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D015544:Inositol 1,4,5-Trisphosphate; D002766:Cholecystokinin; D002784:Cholesterol; D019204:GTP-Binding Proteins; D012844:Sincalide; D005459:Fluorides; C032311:aluminum fluoride