Biliary epithelial cells (BEC) were isolated from normal rat liver with high purity (> 95%) as revealed by morphological criteria as well as staining for gamma-glutamyl transferase and cytokeratin 19. During cultivation for 96 hr flattening of the cells and a loss of microvilli was apparent, while the cytokeratin 19-positive phenotype was maintained. The BEC contained a sodium-dependent as well as a sodium-independent uptake system for glutamate with high capacity. Both activities increased transiently during cultivation peaking after 72 and 48 hr, respectively. After 72 hr, apparent kinetic constants could be calculated for the sodium dependent (Km = 13.6 mM; Vmax = 388 nmoles/min/mg protein) and for the sodium-independent system. (Km = 10.8 mM; Vmax = 132 nmoles/min/mg protein). The transient increase of both transport systems was suppressed by dexamethasone. The sodium-dependence showed a threshold concentration of about 35 mM sodium. Inhibition by kainate was much less potent for BEC than for hepatocytes. These data indicate that BEC contain transport systems for glutamate different from those in hepatocytes and which may be involved in the intrahepatic reabsorbtion of glutamate from bile.
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Abbreviations
- BEC:
-
biliary epithelial cells
- DMEM:
-
Dulbecco's Modified Eagle's Medium
- GGT:
-
gamma-glutamyl transferase
- Dex:
-
dexamethasone
- Glu:
-
glutamate
- N-Me-AIB:
-
N-methyl-aminoisobutyrate
- Hep:
-
hepatocytes
- FBS:
-
Fetal bovine serum
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Eisenmann-Tappe, I., Wizigmann, S. & Gebhardt, R. Glutamate uptake in primary cultures of biliary epithelial cells from normal rat liver. Cell Biol Toxicol 7, 315–325 (1991). https://doi.org/10.1007/BF00124068
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DOI: https://doi.org/10.1007/BF00124068