Wilhelm Roux's archives of developmental biology

, Volume 185, Issue 3, pp 235–248 | Cite as

Retention of the differentiated state by larvalXenopus liver cells in primary culture

  • Walter Wahli
  • Irene Abraham
  • Rudolf Weber
Article

Summary

Electron microscopic analysis of primary cultures derived from larvalXenopus liver has shown that these cells, although they form only two-dimensional aggregates, retain and presumably also develop structural characteristics typical of liver parenchyma cells, such as bile canaliculi with microvilli and epithelial junctional complexes. As judged from structural criteria, primary cultures contain 80–90% hepatocytes. In contrast to the intact tissue, primary cultures showed excessive development of microfilaments, however.

Incorporation of labeled amino acids has revealed further that the capacity for protein synthesis is maintained in culture and that synthesis of liverspecific protein albumin is maintained in vitro, even in liver cultures derived from thyrostatic tadpoles. This latter result suggests that initiation of albumin synthesis in the larval liver is probably not dependent upon thyroid hormones but rather reflects the protodifferentiated state of this tissue.

Key words

Liver Primary culture Ultrastructure Albumin synthesis Xenopus 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Berridge, M.V., Farmer, S.R., Green, C.D., Henshaw, E.C., Tata, J.R.: Characterization of polysomes from Xenopus liver synthesizing vitellogenin and translation of vitellogenin and albumin messenger RNA's in vitro. Eur. J. Biochem.62, 161–172 (1976)Google Scholar
  2. Berridge, M.V., Lane, C.D.: Translation of Xenopus liver messenger RNA in Xenopus oocytes: vitellogenin synthesis and conversion to yolk platelet proteins. Cell8, 283–297 (1976)Google Scholar
  3. Bisbee, C.A., Baker, M.A., Wilson, A.C., Hadzi-Azimi J., Fischberg, M.: Albumin phylogeny for clawed frogs (Xenopus). Science195, 785–787 (1977)Google Scholar
  4. Bragina, E.E., Vasliev, Ju.M., Gelfand, I.M.: Formation of bundles of microfilaments during spreading of fibroblasts on the substrate. Exp. Cell Res.97, 241–248 (1976)Google Scholar
  5. Chen, P.S.: Patterns and metamorphic changes of serum proteins in amphibia. Wilhelm Roux' Archiv165, 132–149 (1970)Google Scholar
  6. Davis, B.J.: Disc electrophoresis. II. Method and application to human serum protein. Ann. N.Y. Acad. Sci.121, 404–427 (1964)Google Scholar
  7. Di Pasquale, A.: Locomotion of epithelial cells. Exp. Cell Res.95, 425–439 (1975)Google Scholar
  8. Fairbanks, G., Stesick, T.L., Wallach, D.F.H.: Electrophoresis analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry10, 2606–2617 (1971)Google Scholar
  9. Frieden, E., Just, J.J.: Hormone responses in amphibian metamorphosis. In: Mechanisms of Hormone Action (G. Litwack, ed.), Vol. 1, pp. 1–52. New York: Academic Press 1970Google Scholar
  10. Goldman, R.D., Lazarides, E., Pollack, R., Weber, K.: The distribution of actin in non-muscle cells. Exp. Cell. Res.90, 333–344 (1975)Google Scholar
  11. Grieninger, G., Granick, S.: Synthesis and differentiation of plasma proteins in cultured embryonic chicken liver cells: A system for study of regulation of protein synthesis. Proc. Natl. Acad. Sci. USA72, 5007–5011 (1975)Google Scholar
  12. Jeejeebhoy, K.N., Ho, J., Greenberg, G.R., Phillips, M.J., Bruce-Robertson, A., Sodtke, U.: Albumin fibrinogen and transferrin synthesis in isolated rat hepatocyte suspensions. Biochem. J.146, 141–155 (1975)Google Scholar
  13. Kistler, A., Weber, R.: A combined biochemical and morphometric study on tissue changes in Xenopus larvae during induced metamorphosis. Mol. cell. Endocrinol.2, 261–288 (1975)Google Scholar
  14. Laemmli, U.K., Favre, M.: Maturation of the head of bacteriophage T4. J. Mol. Biol.80, 575–599 (1973)Google Scholar
  15. Laskey, R.A., Mills, A.D.: Quantitative film detection of3H and14C in polyacrylamide gels by fluorography. Eur. J. Biochem.56, 335–341 (1975)Google Scholar
  16. Lemonnier, F., Cornec, M.L., Golovtchenko, M., Gautier, M.: Synthèse de l'albumine au cours de cultures issues de parenchyme hépatique d'origine humaine. Biochimie56, 1111–1117 (1974)Google Scholar
  17. Ouchterlony, O., Nilsson, L.A.: In: Handbook of Experimental Immunology (D.M. Weir ed.). Oxford Blackwell Scientific 1973Google Scholar
  18. Périssel, B., Chassebeuf, M., Malet, P., Padieu, P.: Etude ultrastructurale d'hépatocytes de rat nouveau-né en culture monocouche établie. C.R. Acad. Sci.277, 2429–2432 (1973)Google Scholar
  19. Reynolds, E.S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol.17, 208–212 (1967)Google Scholar
  20. Rutter, W.J., Wessels, N.K., Grobstein, C.: Control of specific synthesis in the developing pancreas. Nat. Canc. Inst. Monogr. No.13, pp. 51–65 (1964)Google Scholar
  21. Travis, J., Bowen, J., Tewksbury, D., Johnson, D., Pannell, R.: Isolation of albumin from whole human plasma and fractionation of albumin-depleted plasma. Biochem. J.157, 301–306 (1976)Google Scholar
  22. Wahli, W., Weber, R.: Factors promoting the establishment of primary cultures of liver cells from Xenopus larvae. Wilhelm Roux's Archives182, 347–360 (1977)Google Scholar
  23. Wanson, J.-C., Drochmans, P., Mosselmans, R., Ronceveaux, M.F.: Adult rat hepatocytes in primary monolayer culture. Ultrastructural characteristics of intercellular contacts and cell membrane differentiation. J. Cell Biol.74, 858–877 (1977)Google Scholar
  24. Weber, R.: Biochemistry of amphibian metamorphosis. In: The Biochemistry of Animal Development (R. Weber, ed.), Vol. 2, pp. 277–301, New York, Academic Press 1967Google Scholar
  25. Weeke, B.: General Remarks on Principles Equipment, Reagents and Procedures. In: Quantitative Immunoelectrophoresis (N.H. Axelsen, J. Krøll, and B. Weeke, eds.), pp. 15–36 Oslo: Universitetsforlaget 1973Google Scholar
  26. Weigand, K., Müller, M., Urban, J., Schreiber, G.: Intact endoplasmic reticulum and albumin synthesis in rat liver cell suspensions. Exp. Cell Res.67, 27–32 (1971)Google Scholar

Copyright information

© Springer-Verlag 1978

Authors and Affiliations

  • Walter Wahli
    • 1
  • Irene Abraham
    • 1
  • Rudolf Weber
    • 1
  1. 1.Zoological Institute, Division of Cell and Developmental BiologyUniversity of BernBernSwitzerland

Personalised recommendations