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Scanning laser cytometry: alterations induced by cholestatic agents in isolated rat hepatocyte couplets

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Abstract

Scanning laser cytometry, an analytic technique that provides an accurate fluorescent measurement in adherent cells, was used to study cholestatic mechanisms in isolated rat hepatocyte couplets (IRHC). Treatment of IRHC with cholestatic compounds induced a pericanalicular F-actin accumulation and an increase in cytosolic free calcium. These data obtained with a scanning cytometer used in conjunction with anin vitro model representing the primary secretory unit suggest that abnormalities of pericanalicular F-actin filaments and calcium homeostasis play a key role in cholestasis. Considering the necessity for the development of mechanistic studies in toxicology, this technique should prove to be an outstanding tool.

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Abbreviations

ANIT:

α-naphthylisothiocyanate

[Ca2+]i :

cytosolic calcium

CF/TF:

canalicular area fluorescence/total couplet fluorescence

EE:

ethynil estradiol

ERY:

erythromycin estolate

FCS:

fetal calf serum

FITC-phalloidin:

fluorescein isothiocyanate phalloidin

Indo-1-AM:

indo-1-acetyloxymethyl ester

IRHC:

isolated rat hepatocyte couplet

L15:

Leibovitz 15

PH:

phalloidin

TEST:

methyltestosterone

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  1. Institut de Recherches Internationales Servier, 6, Place des Pléiades, 92415, Courbevoie Cédex, France

    N. Thibault

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  1. N. Thibault
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Thibault, N. Scanning laser cytometry: alterations induced by cholestatic agents in isolated rat hepatocyte couplets. Cell Biol Toxicol 10, 323–328 (1994). https://doi.org/10.1007/BF00755778

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  • DOI: https://doi.org/10.1007/BF00755778

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