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Am J Physiol Gastrointest Liver Physiol 278: G251-G258, 2000;
0193-1857/00 $5.00
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Vol. 278, Issue 2, G251-G258, February 2000

Defect of receptor-G protein coupling in human gallbladder with cholesterol stones

Zuo-Liang Xiao, Qian Chen, Joseph Amaral, Piero Biancani, and Jose Behar

Departments of Medicine and Surgery, Rhode Island Hospital and Brown University School of Medicine, Providence, Rhode Island 02903

Human gallbladders with cholesterol stones (ChS) exhibit an impaired muscle contraction and relaxation and a lower CCK receptor-binding capacity compared with those with pigment stones (PS). This study was designed to determine whether there is an abnormal receptor-G protein coupling in human gallbladders with ChS using 35S-labeled guanosine 5'-O-(3-thiotriphosphate) ([35S]GTPgamma S) binding, 125I-labeled CCK-8 autoradiography, immunoblotting, and G protein quantitation. CCK and vasoactive intestinal peptide caused significant increases in [35S]GTPgamma S binding to Galpha i-3 and Gsalpha , respectively. The binding was lower in ChS than in PS (P < 0.01). The reduced [35S]GTPgamma S binding in ChS was normalized after the muscles were treated with cholesterol-free liposomes (P < 0.01). Autoradiography and immunoblots showed a decreased optical density (OD) for CCK receptors, an even lower OD value for receptor-G protein coupling, and a higher OD for uncoupled receptors or Galpha i-3 protein in ChS compared with PS (P < 0.001). G protein quantitation also showed that there were no significant differences in the Galpha i-3 and Gsalpha content in ChS and PS. We conclude that, in addition to an impaired CCK receptor-binding capacity, there is a defect in receptor-G protein coupling in muscle cells from gallbladder with ChS. These changes may be normalized after removal of excess cholesterol from the plasma membrane.

immunoblotting; 35S-labeled guanosine 5'-O-(3-thiotriphosphate) binding; autoradiography; G protein quantitation; smooth muscle.


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