Activation of tyrosine kinase pathway by vanadate in gallbladder smooth muscle.
Vanadate, an inhibitor of tyrosine phosphatase activity, might induce gallbladder contraction through the stimulation of the tyrosine kinase pathway. The aim of this study was to characterize the effects of vanadate in the guinea pig gallbladder smooth muscle. Vanadate exerts contractile effects which are not mediated by neurotransmitter release. The tyrosine kinase inhibitor genistein nearly abolished vanadate contraction, suggesting that an increase in protein tyrosine phosphorylation mediates the actions of vanadate. This suggestion was confirmed by Western blot analysis. Vanadate contractions were reduced in the presence of methoxyverapamil or in Ca(2+)-free medium, suggesting that vanadate may induce Ca(2+) influx. Neither inactivation of the Na(+)/K(+) pump nor reversal of the Na(+)/Ca(2+) exchanger can account for vanadate's actions. Vanadate contractile effects were reduced by indomethacin, as well as mepacrine, the inhibitor of phospholipase A(2), but were not affected by phospholipase C inhibitors. Neither inhibitors of diacylglycerol lipase nor protein kinase C reduced the response induced by vanadate. These data indicate that the effects of vanadate on smooth muscle are mainly mediated by protein tyrosine phosphorylation and reveal a new link between tyrosine phosphorylation and arachidonic acid metabolism in the control of gallbladder smooth muscle contraction.
Alcón
S
S
Department of Physiology, University of Extremadura, 10071, Cáceres, Spain.
Camello
P J
PJ
García
L J
LJ
Pozo
M J
MJ
D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D004789:Enzyme Activation / Q000187:drug effects; D005704:Gallbladder / Q000187:drug effects / Q000502:physiology; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects*; D009130:Muscle, Smooth / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D018377:Neurotransmitter Agents / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology*
D018377:Neurotransmitter Agents; D019000:Phosphotyrosine; D014638:Vanadates; D011505:Protein-Tyrosine Kinases; D002118:Calcium