In vivo dynamic metabolic imaging of obstructive cholestasis in mice
Journal
American Journal of Physiology - Gastrointestinal and Liver Physiology
Journal Volume
296
Journal Issue
5
Pages
G1091-G1097
Date Issued
2009
Author(s)
Abstract
We tried to image obstructive cholestasis by using a newly developed imaging system to measure the alterations of hepatobiliary function in living mice with their bile ducts ligated. A hepatic imaging window was installed on the upper abdomen soon after the mice underwent ligation of the common bile duct. On the next day, the mice received intravenous injection of rhodamine B isothiocyanate-dextran and carboxyfluorescein diacetate. The later would be transformed into fluorogenic carboxyfluorescein (detected at ∼500-550 nm) by hepatocytes and then excreted into bile canaliculi. The images were acquired by multiphoton microscopy. The fluorescence intensities at ∼500-550 nm within hepatocytes or sinusoids were measured in time series. In mice with bile duct ligation, bile canaliculi failed to appear during the whole observation period over 100 min following carboxyfluorescein diacetate injection, whereas the fluorescence was retained much longer within sinusoids. Furthermore, the fluorescence intensities in sinusoids were persistently higher than in hepatocytes during the course. Bile duct ligation impedes hepatocytes to excrete carboxyfluorescein into bile canaliculi. The kinetics of fluorescence intensities in hepatocytes and sinusoids indicated there is an active machinery operating backflow of this fluorogenic bile solute from hepatocytes into sinusoids in the liver with obstructive cholestasis. Copyright © 2009 the American Physiological Society.
Subjects
Common bile duct ligation; Fluorescence imaging; Hepatocyte transporter; Multiphoton microscopy
Other Subjects
carboxyfluorescein diacetate succinimidyl ester; dextran derivative; isothiocyanic acid derivative; rhodamine B; rhodamine B isothiocyanate dextran; unclassified drug; carboxyfluoresceindiacetate; fluorescein derivative; fluorescent dye; multidrug resistance protein; organic anion transporter; rhodamine; rhodamine isothiocyanate; animal experiment; animal model; animal tissue; article; cholestasis; common bile duct; controlled study; fluorescence; image analysis; imaging system; liver cell; liver sinusoid; molecular imaging; mouse; multiphoton microscopy; nonhuman; obstructive cholestasis; priority journal; animal; bile; C57BL mouse; common bile duct; cystic duct; disease model; intrahepatic bile duct; intravenous drug administration; ligation; liver cell; male; metabolism; methodology; pathology; transport at the cellular level; Animals; Bile; Bile Canaliculi; Biological Transport; Cholestasis; Common Bile Duct; Cystic Duct; Disease Models, Animal; Fluoresceins; Fluorescent Dyes; Hepatocytes; Injections, Intravenous; Ligation; Male; Mice; Mice, Inbred C57BL; Microscopy, Fluorescence, Multiphoton; Multidrug Resistance-Associated Proteins; Organic Anion Transporters; Rhodamines
Type
journal article