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Laboratoire de Physiologie Cellulaire et Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Enseignement Supérieur Associé 5017, Université de Bordeaux II, 33076 Bordeaux Cedex, France
In rat portal vein myocytes, Ca2+ signals can be generated by inositol 1,4,5-trisphosphate (InsP3)- and ryanodine-sensitive Ca2+ release channels, which are located on the same intracellular store. Using a laser scanning confocal microscope associated with the patch-clamp technique, we showed that propagated Ca2+ waves evoked by norepinephrine (in the continuous presence of oxodipine) were completely blocked after internal application of an anti-InsP3 receptor antibody. These propagated Ca2+ waves were also reduced by ~50% and transformed in homogenous Ca2+ responses after application of an anti-ryanodine receptor antibody or ryanodine. All-or-none Ca2+ waves obtained with increasing concentrations of norepinephrine were transformed in a dose-response relationship with a Hill coefficient close to unity after ryanodine receptor inhibition. Similar effects of the ryanodine receptor inhibition were observed on the norepinephrine- and ACh-induced Ca2+ responses in non-voltage-clamped portal vein and duodenal myocytes and on the norepinephrine-induced contraction. Taken together, these results show that ryanodine-sensitive Ca2+ release channels are responsible for the fast propagation of Ca2+ responses evoked by various neurotransmitters producing InsP3 in vascular and visceral myocytes.
inositol 1,4,5-trisphosphate receptors; cytosolic calcium; confocal microscopy
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