• actin;
  • calcium ionophore;
  • calmodulin;
  • cholestasis;
  • fura 2;
  • microfluorometry

ABSTRACT— Dynamic contractions of bile canaliculi have been observed in cultured doublet hepatocytes by means of time-lapse cinephotomicrography, and this contractile movement plays an important role in bile secretion. Although details of the mechanism are still unknown, the Ca2+-calmodulin system is believed to play a main role in this mechanism. In this study we measured the intracellular Ca2+ concentration of individual doublet hepatocytes using the Ca2+ indicator “fura 2” and microscopic fluorometry. We also observed the effects of A23187, norepinephrine and epinephrine on bile canalicular contraction and intracellular Ca2+ concentration. After loading 1 μmol/1 fura 2 in doublet cells, we added A23187, epinephrine or norepinephrine and then measured the Ca2+ concentration in a given small area in the cytoplasm of individual doublet cell. A23187, norepinephrine and epinephrine caused a prompt increase of the intracellular Ca2+ concentration and also caused bile canalicular contraction. The present study indicates that the sudden increase of intracellular Ca2+ concentration causes bile canalicular contraction through the Ca2+-calmodulin system.