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. 1983 Sep;3(9):1552–1561. doi: 10.1128/mcb.3.9.1552

Changes in liver-specific compared to common gene transcription during primary culture of mouse hepatocytes.

D F Clayton, J E Darnell Jr
PMCID: PMC370008  PMID: 6633533

Abstract

Liver-specific mRNA sequences were examined in primary cultures of mouse hepatocytes. After cell disaggregation by collagenase treatment and for at least 24 h in culture, little change in liver-specific mRNA concentrations was noted. Gradually over a period of 140 h, liver-specific mRNAs declined. In contrast, transcriptional assays in which liver cell nuclei were used to produce 32P-labeled nuclear RNA showed that liver-specific gene transcription was greatly diminished within 24 h, while polymerase II transcription of "common" genes and transcription of tRNA and rRNA did not decline. Thus, a prompt differential transcriptional effect seems to underlie the gradual loss of tissue specificity of the primary cultures.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ben-Ze'ev A., Farmer S. R., Penman S. Protein synthesis requires cell-surface contact while nuclear events respond to cell shape in anchorage-dependent fibroblasts. Cell. 1980 Sep;21(2):365–372. doi: 10.1016/0092-8674(80)90473-0. [DOI] [PubMed] [Google Scholar]
  2. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  3. Derman E., Krauter K., Walling L., Weinberger C., Ray M., Darnell J. E., Jr Transcriptional control in the production of liver-specific mRNAs. Cell. 1981 Mar;23(3):731–739. doi: 10.1016/0092-8674(81)90436-0. [DOI] [PubMed] [Google Scholar]
  4. Durnam D. M., Palmiter R. D. Transcriptional regulation of the mouse metallothionein-I gene by heavy metals. J Biol Chem. 1981 Jun 10;256(11):5712–5716. [PubMed] [Google Scholar]
  5. Fukuda-Taira S. Hepatic induction in the avian embryo: specificity of reactive endoderm and inductive mesoderm. J Embryol Exp Morphol. 1981 Jun;63:111–125. [PubMed] [Google Scholar]
  6. Gallin W. J., Edelman G. M., Cunningham B. A. Characterization of L-CAM, a major cell adhesion molecule from embryonic liver cells. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1038–1042. doi: 10.1073/pnas.80.4.1038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Greengard O. Enzymic differentiation in mammalian liver injection of fetal rats with hormones causes the premature formation of liver enzymes. Science. 1969 Feb 28;163(3870):891–895. doi: 10.1126/science.163.3870.891. [DOI] [PubMed] [Google Scholar]
  8. Greengard O., Federman M., Knox W. E. Cytomorphometry of developing rat liver and its application to enzymic differentiation. J Cell Biol. 1972 Feb;52(2):261–272. doi: 10.1083/jcb.52.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Harpold M. M., Evans R. M., Salditt-Georgieff M., Darnell J. E. Production of mRNA in Chinese hamster cells: relationship of the rate of synthesis to the cytoplasmic concentration of nine specific mRNA sequences. Cell. 1979 Aug;17(4):1025–1035. doi: 10.1016/0092-8674(79)90341-6. [DOI] [PubMed] [Google Scholar]
  10. Hofer E., Darnell J. E., Jr The primary transcription unit of the mouse beta-major globin gene. Cell. 1981 Feb;23(2):585–593. doi: 10.1016/0092-8674(81)90154-9. [DOI] [PubMed] [Google Scholar]
  11. Houssaint E. Differentiation of the mouse hepatic primordium. I. An analysis of tissue interactions in hepatocyte differentiation. Cell Differ. 1980 Oct;9(5):269–279. doi: 10.1016/0045-6039(80)90026-3. [DOI] [PubMed] [Google Scholar]
  12. Kafatos F. C., Jones C. W., Efstratiadis A. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 1979 Nov 24;7(6):1541–1552. doi: 10.1093/nar/7.6.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kioussis D., Eiferman F., van de Rijn P., Gorin M. B., Ingram R. S., Tilghman S. M. The evolution of alpha-fetoprotein and albumin. II. The structures of the alpha-fetoprotein and albumin genes in the mouse. J Biol Chem. 1981 Feb 25;256(4):1960–1967. [PubMed] [Google Scholar]
  14. Klaunig J. E., Goldblatt P. J., Hinton D. E., Lipsky M. M., Chacko J., Trump B. F. Mouse liver cell culture. I. Hepatocyte isolation. In Vitro. 1981 Oct;17(10):913–925. doi: 10.1007/BF02618288. [DOI] [PubMed] [Google Scholar]
  15. Klaunig J. E., Goldblatt P. J., Hinton D. E., Lipsky M. M., Trump B. F. Mouse liver cell culture. II. Primary culture. In Vitro. 1981 Oct;17(10):926–934. doi: 10.1007/BF02618289. [DOI] [PubMed] [Google Scholar]
  16. Leffert H. L., Moran T., Boorstein R., Koch K. S. Procarcinogen activation and hormonal control of cell proliferation in differentiated primary adult rat liver cell cultures. Nature. 1977 May 5;267(5606):58–61. doi: 10.1038/267058a0. [DOI] [PubMed] [Google Scholar]
  17. Leffert H., Moran T., Sell S., Skelly H., Ibsen K., Mueller M., Arias I. Growth state-dependent phenotypes of adult hepatocytes in primary monolayer culture. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1834–1838. doi: 10.1073/pnas.75.4.1834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Michalopoulos G., Pitot H. C. Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp Cell Res. 1975 Aug;94(1):70–78. doi: 10.1016/0014-4827(75)90532-7. [DOI] [PubMed] [Google Scholar]
  19. Newman S., Guzelian P. S. Stimulation of de novo synthesis of cytochrome P-450 by phenobarbital in primary nonproliferating cultures of adult rat hepatocytes. Proc Natl Acad Sci U S A. 1982 May;79(9):2922–2926. doi: 10.1073/pnas.79.9.2922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pitot H. C., Sirica A. E. Methodology and utility of primary cultures of hepatocytes from experimental animals. Methods Cell Biol. 1980;21B:441–456. doi: 10.1016/s0091-679x(08)60697-4. [DOI] [PubMed] [Google Scholar]
  21. Salditt-Georgieff M., Harpold M., Sawicki S., Nevins J., Darnell J. E., Jr Addition of poly(A) to nuclear RNA occurs soon after RNA synthesis. J Cell Biol. 1980 Sep;86(3):844–848. doi: 10.1083/jcb.86.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wiskocil R., Bensky P., Dower W., Goldberger R. F., Gordon J. I., Deeley R. G. Coordinate regulation of two estrogen-dependent genes in avian liver. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4474–4478. doi: 10.1073/pnas.77.8.4474. [DOI] [PMC free article] [PubMed] [Google Scholar]

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