Skip to main content
Log in

Amino acid transport in established adult rat liver epithelial cell lines

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

The capacities of Na+-dependent transport of α-aminoisobutyrate, glutamine and glutamate in four established and three transformed rat liver epithelial cell lines were found to be considerably higher than those of isolated and cultured hepatocytes. At least for transport systems A and G this seemed to be due to elevated values of V max , whereas the values for K m were quite comparable to those of hepatocytes. In contrast to hepatocytes, however, no significant hormonal stimulation of amino acid uptake could be detected in the cell lines.

Each normal cell line expressed a distinct pattern of transport capacities with respect to the three systems measured and this was not altered by chemical transformation of the lines. The individual patterns of the lines showed no similarity to presumptive patterns of subpopulations of liver parenchymal cells. In particular, there was no evidence for a direct relationship of one of the cell lines with a small subpopulation of parenchymal cells located adjacent to hepatic venules as revealed by additional measurements of glutamine synthetase, a marker enzyme for this particular subpopulation.

It is concluded that established rat liver epithelial cell lines express features characteristic of normal hepatocytes with respect to amino acid transport, but have developed a distinct phenotype adapted to a rapid, hormone-independent growth in vitro. Alteration of their phenotype by transformation is not coupled with a further increase in amino acid transport capacity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

AIB:

α-aminoisobutyrate

LPC:

liver parenchymal cells

N-Methyl-AIB:

N-methyl-α-aminoisobutyrate

TAT:

tyrosine aminotransferase

References

  • BISSELL, M.D. (1976). Study of hepatocyte function in cell culture. Prog. Liver. Dis. 5:69–82.

    Google Scholar 

  • CHESSEBEUF, M., OLSSON, A., BOURNOT, P., DESGRES, J., GUIGUET, M., MAUME, G., MAUME, B.F., PERISSEL, B. and PADIEU, P. (1975). Retention and loss of certain enzymes in various primary cultures and cell lines of normal rat liver. In: Gene Expression and Carcinogenesis in Cultured Liver (L.E. Gerschenson and B.E. Thompson, eds.) pp. 94–118. Academic Press, New York.

    Google Scholar 

  • CHESSEBEUF, M., FISCHBACH, M. and PADIEU, P. (1984). Time course study of L-tyrosine aminotransferase induction in rat liver cell lines. Cell Biol. Toxicol. 1:31–40.

    Google Scholar 

  • CHRISTENSEN, H.N. (1984). Organic anion transport during seven decades. The amino acids. Biochim. Biophys. Acta 779:255–269.

    Google Scholar 

  • CHRISTENSEN, H.N., MAKOWSKE, M. and VADGAMA, J.V. (1983). Changes of amino acid transport systems on maturation and transformation of rat hepatocytes. How can ASC serve for amino acids of unlike charge? In: Isolation, Characterization and Use of Hepatocytes (R.A. Harris and N.W. Cornell, eds.) pp. 217–226. Elsevier Biomedical, New York.

    Google Scholar 

  • EAGLE, H., PIEZ, K. A. and LEVI, M. (1961). The intracellular amino acid concentrations required for protein synthesis in cultured human cells. J. Biol. Chem. 236:2039–2042.

    Google Scholar 

  • FOSTER, D.O. and PARDEE, A.B. (1969). Transport of amino acids by confluent and nonconfluent 3T3 and polyoma virus-transformed 3T3 cells growing on glass cover slips. J. Biol. Chem. 244:2675–2681.

    Google Scholar 

  • FURUKAWA, K., SHIMADA, T., ENGLAND, P., MOCHIZUKI, Y. and WILLIAMS, G. M. (1986). Enrichment and characterization of clonigenic cells from adult rat liver and a technique for consistent development of epithelial cell lines. In Vitro, submitted for publication.

  • GAZZOLA, G. C., DALL ASTA, V., FRANCHI-GAZZOLA, R. and WHITE, M.F. (1981). The cluster-tray method for rapid measurement of solute fluxes in adherent cultured cells. Anal. Biochem. 115:368–374.

    Google Scholar 

  • GEBHARDT, R. and MECKE, D. (1979).Perifused monolayer cultures of rat hepatocytes as an improved in vitro system for studies on ureogenesis. Exp. Cell Res. 124:349–359.

    Google Scholar 

  • GEBHARDT, R. and MECKE, D. (1983). Glutamate uptake by cultured rat hepatocytes is mediated by hormonally inducible, sodium-dependent transport systems. FEBS Lett. 161:275–278.

    Google Scholar 

  • GEBHARDT, R. and MECKE, D. (1983b). Heterogeneous distribution of glutamine synthetase among rat liver parenchymal cells in situ and in primary culture. EMBO J. 2:567–570.

    Google Scholar 

  • GEBHARDT, R. and MECKE, D. (1984a). Cellular distribution and regulation of glutamine synthetase in liver. In Glutamine Metabolism in Mammalian Tissues (D. Haussinger and H. Sies, eds.) pp. 98–121. Springer Verlag, Berlin, Heidelberg, New York.

    Google Scholar 

  • GEBHARDT, R. and MECKE, D. (1984b). Intercellular compartmentation of hepatic ammonia metabolism: A basis for the differential regulatory influence of hormones on related enzymes and amino acid transport systems. In: Advances in Hepatic Encephalopathy and Urea Cycle Diseases (G. Kleinberger, P. Ferency, P. Riederer and H. Thaler, eds.) pp. 126–131. Karger, Basel.

    Google Scholar 

  • GEBHARDT, R. (1986). Hormonal regulation of glutamate transport in cultured rat hepatocytes. Submitted for publication.

  • GERSCHENSON, L.E., DAVIDSON, M.B. and ANDERSSON, M. (1974). Hormonal regulation of rat liver cells cultured in chemically defined medium. Eur. J. Biochem. 41:139–148.

    Google Scholar 

  • GRISHAM, J.W., THAL, S.B. and NAGEL, A. (1975). Cellular derivation of continuously cultured epithelial cells from normal rat liver. In: Gene Expression and Carcinogenesis in Cultured Liver (L.E. Gerschenson and B.E. Thompson, eds.) pp. 1–23. Academic Press, New York.

    Google Scholar 

  • ISSELSBACHER, K.J. (1972). Increased uptake of amino acids and 2-deoxy-D-glucose by virus transformed cells in culture. Proc. Natl. Acad. Sci. USA 69:585–589.

    Google Scholar 

  • KILBERG, M.S. (1982). Amino acid transport in isolated rat hepatocytes. J. Membrane Biol. 69:1–12.

    Google Scholar 

  • KILBERG, M.S., HANDLOGTEN, M.E. and CHRISTENSEN, H.N. (1980). Characteristics of an amino acid transport system in rat liver for glutamine, asparagine, histidine, and closely related analogs. J. Biol. Chem. 255:4011–4019.

    Google Scholar 

  • KOCH, M.R. and LEA, M.A. (1981). Uptake of 14C-labeled dicarboxylic amino acids in hepatocytes and hepatoma cells. Cancer Research 41:3065–3070.

    Google Scholar 

  • LE CAM, A., REY, J.-F., FEHLMANN, M., KITABGI, P. and FREYCHET, P. (1979). Amino acid transport in isolated hepatocytes after partial hepatectomy in the rat. Am. J. Physiol. 5:E594-E602.

    Google Scholar 

  • LEVINTOW, L. (1954). The glutamyltransferase activity of normal and neoplastic tissues. J. Natl. Cancer 15:347–352.

    Google Scholar 

  • MALAN-SHIBLEY, L. and IYPE, P.T. (1981). The influence of culture conditions on cell morphology and tyrosine aminotransferase levels in rat liver epithelial cell lines. Exp. Cell Res. 131:363–371.

    Google Scholar 

  • MAYER, D. and SCHÄFER, B. (1982). Biochemical and morphological characterization of glycogen-storing epithelial liver cell lines. Exp. Cell Res. 138:1–14.

    Google Scholar 

  • SHIMADA, T., FURUKAWA, K., KREISER, D.M., CAWEIN, A. and WILLIAMS, G. M. (1983). Induction of transformation by six classes of chemical carcinogens in adult rat liver epithelial cells. Cancer Res. 43:5087–5092.

    Google Scholar 

  • SHORT, J., ARMSTRONG, N.B., ZEMEL, R. and LIEBERMAN, I. (1973). A role for amino acids in the induction of deoxyribonucleic acid synthesis in liver. Biochem. Biophys. Res. Commun. 50:430–437.

    Google Scholar 

  • SHOTWELL, M.A., KILBERG, M.S. and OXENDER, D.L. (1983). The regulation of neutral amino acid treatment in mammalian cells. Biochem. Biophys. Acta 737:267–284.

    Google Scholar 

  • SIES, H. and HÄUSSINGER, D. (1984). Hepatic glutamine and ammonia metabolism. In: Glutamine Metabolism in Mammalian Tissues (D. Haussinger, and H. Sies, eds.) pp. 78–97. Springer Verlag, Heidelberg, New York.

    Google Scholar 

  • TSAO, M.-S., SMITH, J.D., NELSON, K.G. and GRISHAM, J.W. (1984). A diploid epithelial cell line from normal adult rat liver with phenotypic properties of "Oval" cells. Exp. Cell. Res. 154:38–52.

    Google Scholar 

  • VADGAMA, J.V. and CHRISTENSEN, H.N. (1983). Comparison of system N in fetal hepatocytes and in related cell lines. J. Biol. Chem. 258:6422–6429.

    Google Scholar 

  • WILLIAMS, G. M. (1975). The study of chemical carcinogenesis using cultured liver cells. In: Gene Expression and Carcinogenesis in Cultured Liver (L.E. Gerschenson and B.E. Thompson, eds.) pp. 480–487. Academic Press, New York.

    Google Scholar 

  • WILLIAMS, G.M. (1976). Primary and long-term culture of rat liver epithelial cells. Methods Cell Biol. 14:357–364.

    Google Scholar 

  • WILLIAMS, G.M., BERMUDEZ, E. and SCARAMUZZINO, D. (1977). Rat hepatocyte primary cell cultures. III. Improved dissociation and attachment techniques and the enhancement of survival by culture medium. In Vitro 13:809–817.

    Google Scholar 

  • WILLIAMS, G.M., STROMBERG, K. and KROES, R. (1973). Cytochemical and ultrastructural alterations associated with confluent growth in cell cultures of epithelial-like cells from rat liver. Lab. Invest. 29:293–303.

    Google Scholar 

  • WILLIAMS, G.M. and GUNN, J.M. (1974). Long-term cell culture of adult rat liver epithelial cells. Exp. Cell Res. 89:139–142.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gebhardt, R., Williams, G.M. Amino acid transport in established adult rat liver epithelial cell lines. Cell Biol Toxicol 2, 9–20 (1986). https://doi.org/10.1007/BF00117703

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00117703

Keywords

Navigation