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Am J Physiol Gastrointest Liver Physiol 274: G283-G289, 1998;
0193-1857/98 $5.00
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Vol. 274, Issue 2, G283-G289, February 1998

Impaired G protein function in gallbladder muscle from progesterone-treated guinea pigs

Qian Chen, Vikas Chitinavis, Zouliang Xiao, Peirong Yu, Sangik Oh, Piero Biancani, and Jose Behar

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

This study was designed to elucidate the mechanism of action of progesterone on gallbladder smooth muscle in guinea pigs. Adult male guinea pigs were treated with either progesterone (2 mg · kg-1 · day-1) or saline for 7 days. Gallbladder muscle cells were isolated by enzymatic digestion with collagenase. Contractile responses to agonists were expressed as percent shortening from control cell length. [35S]guanosine 5'-O-(3-thiotriphosphate) ([35S]GTPgamma S)-binding properties of G proteins were assessed in crude membranes of gallbladder muscle with or without cholecystokinin octapeptide (CCK-8) stimulation. Gallbladder muscle cells from progesterone-treated guinea pigs exhibited an impaired contractile response to CCK-8, GTPgamma S, or aluminum fluoride but a normal response to potassium chloride or D-myo-inositol 1,4,5-trisphosphate compared with controls. Western blot analysis of gallbladder muscle revealed the presence of Gi 1-2, Gi 3, Gq/11, and Gs proteins. The maximal contraction induced by CCK-8 was blocked by pertussis toxin and Gialpha 3-specific antibodies, but not by Gialpha 1-2 or Gq/11alpha antibodies. CCK-8 caused a significant increase in [35S]GTPgamma S binding to Gialpha 3, but not to Gq/11alpha or Gialpha 1-2. The stimulation of Gialpha 3 binding, however, was significantly reduced in gallbladder muscle membranes from progesterone-treated guinea pigs compared with that in control animals. In conclusion, progesterone might cause gallbladder hypomotility by downregulating Gi 3 proteins.

cholecystokinin; smooth muscle


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