PMID: 9193924May 1, 1997Paper

Canalicular retention as an in vitro assay of tight junctional permeability in isolated hepatocyte couplets: effects of protein kinase modulation and cholestatic agents

Fundamental and Applied Toxicology : Official Journal of the Society of Toxicology
M G RomaR Coleman

Abstract

A simple, fast method to evaluate acute changes of tight junctional permeability in isolated hepatocyte couplets is proposed. The method consists of the recording of the number of canalicular vacuoles able to retain the previously accumulated fluorescent bile acid analogue cholyl-lysyl-fluorescein (CLF), as visualized by inverted fluorescent microscopy, following acute exposure to the compounds under study. The method was validated by (i) making a systematic documentation of the effect on CLF retention of a variety of hormonal modulators (vasopressin and phorbol esters), as well as several cholestatic (taurolithocholic acid, cyclosporin A, and estradiol 17 beta-glucuronide) and hepatotoxic agents (menadione, A23187, and t-butyl hydroperoxide), all known to affect biliary permeability in intact liver, and (ii) carrying out a comparative analysis of the results obtained with those recorded using rapid canalicular access of horseradish peroxidase (HRP) as an alternative procedure. The compounds tested all decreased canalicular vacuolar retention of CLF in a dose-dependent manner. Vasopressin- and phorbol ester-induced decline in CLF retention were prevented by pretreatment with the protein kinase C inhibitors H-7 and staurosporine...Continue Reading

Citations

Oct 1, 2015·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Michaël MaesMathieu Vinken
Oct 2, 1998·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·M G RomaR Coleman
Oct 31, 2008·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Fernando A CrocenziMary Vore
Jul 1, 2006·Critical Reviews in Toxicology·Mathieu VinkenTamara Vanhaecke
Feb 2, 2013·Drug Metabolism Reviews·Eva RamboerMathieu Vinken
Oct 17, 1998·Toxicology and Applied Pharmacology·J Ahmed-ChoudhuryR Coleman
Jun 3, 2021·International Journal of Molecular Sciences·Andrés TabernillaMathieu Vinken

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