Virchows Archiv B

, Volume 55, Issue 1, pp 57–64 | Cite as

Nuclear growth and chromatin relaxation-condensation cycle in hepatocytes during the proliferative activation of rat liver

  • Joan Serratosa
  • Jordi Domingo
  • Carles Enrich
  • Oriol Bachs


In order to quantify the changes in nucleolar and nuclear volumes and in chromatin condensation produced during proliferative activation we have carried out morphometric studies on hepatocyte nuclei during rat liver regeneration using electron microscopy. To minimize the artefactual effects produced by fixation on subcellular structures we have fixed the livers by perfusion with gluteraldehyde. The mean values for the nucleolar and nuclear volumes were progressively increased until 28 h after 66% partial hepatectomy. The maximum values raised for the nuclei and nucleoli at this time were 3 and 4.28 times, respectively, those of controls. Later, nuclear and nucleolar volumes progressively declined. Two waves of diminution in nuclear electron-dense material were produced after hepatectomy. The first occurred between 0 and 12 h, with minimum values 1.34 times lower than those from control animals at 8 h. The second occurred between 12 and 28 h, with minimum values 2.56 times lower than those from control rats at 24 h. These two waves in chromatin relaxation correlate very well with the transcriptional changes described by other authors during the pre-replicative, replicative and mitotic phases of liver regeneration.

Key words

Liver regeneration Nucleus Chromatin Morphometry Cell cycle Rat 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abercrombie M (1946) Stimation of nuclear population from microtomic sections. Anat Rec 94:239–247CrossRefGoogle Scholar
  2. Bade EG (1967) Biological and ultrastructural aspects of early stages in liver regeneration. In: Their H, Rytoma T (eds) Control of cellular growth in adult organisms. Academic Press, New York, pp 260–269Google Scholar
  3. Bader G (1969) Elektronmikroskopische Untersuchungen zur funktioneilen Morphologie der regenierenden Liberzelle nach partieller Hepatektomie. Acta Hepatol-Splenol 16:281–297Google Scholar
  4. Becker FF (1969) Cell division in normal mammalian tissues. Ann Rev Med 20:243–258PubMedCrossRefGoogle Scholar
  5. Becker FF (1970) Normal hepatocyte in division: Regeneration of mammalian liver. Progr Liver Dis 3:60–76PubMedGoogle Scholar
  6. Becker FF, Lane BP (1965) Regeneration of mammalian liver. I. Autophagocytosis during dedifferentiation of the liver cell in preparation from cell division. Am J Pathol 47:782–801Google Scholar
  7. Becker FF, Lane BP (1966) Regeneration of mammalian liver. IV. Evidence of role of cytoplasmic alterations in preparation for mitosis. Am J Pathol 49:227–235PubMedGoogle Scholar
  8. Becker FF, Lane BP (1968) Regeneration of mammalian liver. VI. Retention of phenobarbital-induced cytoplasmic alterations in dividing hepatocytes. Am J Pathol 52:211–225PubMedGoogle Scholar
  9. Briane D, Jalozot R, Gontcharoff M (1978) Variation du taux d’euchromatine au cours de la regeneration du foi du rat. Biologie Cellulaire 31:1–6Google Scholar
  10. Brues AM, Drury DR, Brues MC (1936) Quantitative study of cell growth in regenerating liver. Arch Pathol (Chicago) 22:658–673Google Scholar
  11. Bucher NLR (1963) Regeneration of mammalian liver. Int Rev Cytol 15:245–300PubMedCrossRefGoogle Scholar
  12. Bucher NLR, Swaffield MN (1964) The rate of incorporation of labeled thymidine into deoxyribonucleic acid of regenerating rat liver in relation to the amount of liver excised. Cancer Res 24:1611–1625PubMedGoogle Scholar
  13. Bucher NLR, Malt RA (1971) Regeneration of liver and kidney. In: Ingelfinger FJ (ed) New England J Med medical progress series. Little Brown and Company, Boston, pp 28–43Google Scholar
  14. Busch H, Smetana K (1970) The nucleolus. Academic Press, New York, pp 75–82Google Scholar
  15. Chaudhuri S, Lieberman I (1968) Control of ribosome synthesis in normal and regenerating liver. J Biol Chem 243:29–33PubMedGoogle Scholar
  16. Dabeva D, Dudov P (1982) Transcriptional control of ribosome production in regeneration rat liver. Biochem J 208:101–108PubMedGoogle Scholar
  17. Darzynkiewicz Z, Traganos F, Melamed MR (1980) New cell cicle compartments identified by multiparameter flow citometry. Cytochemistry 2:98–108Google Scholar
  18. Derenzini M, Lorenzini E, Marinozzi V, Barotti P (1977) Ultrastructural cytochemistry of active chromatin in regeneratin rat hepatocytes. J Ultrastr Res 59:250–262CrossRefGoogle Scholar
  19. Derenzini M, Pession-Brizzi A, Novello F (1980) Chromatin condensation in isolated rat hepatocyte nuclei by ribonuclease treatment. Experientia 36:181–183PubMedCrossRefGoogle Scholar
  20. Derenzini M, Pession-Brizzi A, Betts-Eusebi C, Novello F (1981) Relationship between the fine structural organization of chromatin and nucleic acid synthesis in regenerating rat hepatocytes. J Ultrastr Res 75:229–242CrossRefGoogle Scholar
  21. Dudov P, Dabeva D (1983) Post-transcriptional regulation of ribosome formation in the nucleus of regenerating rat liver. Biochem J 210:183–192PubMedGoogle Scholar
  22. Fahimi HD (1967) Perfusion and inmersion fixation of rat liver with glutaraldehyde. Lab Invest 16:736–750PubMedGoogle Scholar
  23. Farquhar MG, Palade GE (1965) Cell junctions in amphibian skin. J Cell Biol 26:263–291PubMedCrossRefGoogle Scholar
  24. Fausto N (1984) Messenger RNA in regenerating liver: implications for the understending of regulated growth. Mol Cell Biochem 59:131–147PubMedCrossRefGoogle Scholar
  25. Fausto N (1986) New perspectives on liver regeneration. Hepatology 6:326–327PubMedCrossRefGoogle Scholar
  26. Fawcett DW (1955) Observations on cytology and electron microscopy of hepatic cells. J Nat Cancer Inst 15:1475–1502PubMedGoogle Scholar
  27. Fisher ER, Fisher B (1963) Ultrastructural hepatic changes following partial hepatectomy and portacaval shunt in rat. Lab Invest 12:929–942PubMedGoogle Scholar
  28. Fujioka M, Koga M, Lieberman J (1963) Metabolism of ribonucleic acid after partial hepatectomy. J Biol Chem 238:3401–3406PubMedGoogle Scholar
  29. Fullman RL (1953) Measurement of particle size in opaque bodies. J Metals 5:447–461Google Scholar
  30. Gaub J (1978) Cytophotometric determination of nuclear nonhistone protein in the replicative phase of rat liver regeneration. Exp Cell Res 114:167–174PubMedCrossRefGoogle Scholar
  31. Grisham JW (1962) Morphologyc study of deoxyribonucleic acid synthesis and cell proliferation in regenerating rat liver: Autoradiography with thymidine-3H. Cancer Res 22:842–849PubMedGoogle Scholar
  32. Grisham JW, Tillman MS, Nagel AEH, Compagno MD (1975) Ultrastructure of the proliferating hepatocyte: sinusoidal and endoplasmic reticulum. In: Lesch R, Reutter W (eds) Liver regeneration after experymental injury. Stratton Intercontinental Medical Book Corporation, New York, pp 6–25Google Scholar
  33. Harris H (1968) Nucleus and cytoplasm (1st ed). Clarendom Press, OxfordGoogle Scholar
  34. Herdson PB, Garvin PJ, Jennings RB (1964) Fine structural changes produced in rat liver by partial starvation. Am J Pathol 45:157–181PubMedGoogle Scholar
  35. Higashi K, Matsuhisa Y, Kitao A, Sakamoto Y (1968) Selective suppresion of nucleolar RNA metabolism in absence of protein synthesis. Biochim Biophys Acta 166:388–393PubMedGoogle Scholar
  36. Higgins GM, Anderson RM (1931) Experymental pathology of the liver. I. Restoration of the liver of the white rat following partial surgical removal. Arch Pathol 12:186–202Google Scholar
  37. Jordan SW (1964) Electron microscopy of hepatic regeneration. Exp Mol Pathol 3:183–200CrossRefGoogle Scholar
  38. Lane BP, Becker FF (1966) Regeneration of the mammalian liver. II. Surface alterations during dedifferentiation of the liver cell in preparation for cell division. Am J Pathol 48:183–196PubMedGoogle Scholar
  39. Leffert HL, Koch KS, Moran T, Rubalcava B (1979) Hormonal control of rat liver regeneration. Gastroenterology 76:1470–1482PubMedGoogle Scholar
  40. Lindberg LG, Vorwerk P (1972) Stimation of volumes of ellipsoid bodies in conglomerates through random plane sections. Lab Invst 27:384–386Google Scholar
  41. Lloyd D, Poole RK, Edwards SW (1982) The cell division Cycle. Temporal organitzation and control of cellular growth and reproduction. Academic Press, London New York, pp 143–180Google Scholar
  42. Loud AV (1968) A quantitative stereological description of the ultrastructure of normal rat liver parenchymal cells. J Cell Biol 37:27–46PubMedCrossRefGoogle Scholar
  43. Mironescu S, Dragomir C (1967) Nucleolar behavior in regenerating liver of normal and whole-body-irradiated rats. Cancer Res 27:1819–1830PubMedGoogle Scholar
  44. Mitchison JM (1973) The biology of the cell cycle. Cambridge University Press, pp 128–159Google Scholar
  45. Morin-Coutu MS, Bilodeaw BS, Simpkins H (1977) Structural and compositional changes in rat liver ribosomes following partial hepatectomy. Lab Invest 37:307–313PubMedGoogle Scholar
  46. Muramatsu M, Busch H (1965) Studies on nuclear and nucleolar ribonucleic acid of regenerating rat liver. J Biol Chem 240:3960–3966PubMedGoogle Scholar
  47. Murray AB, Strecker W, Silz S (1981) Ultrastructural changes in rat hepatocytes after partial hepatectomy and comparison with biochemical results. J Cell Sci 50:433–448PubMedGoogle Scholar
  48. Nadal C (1975) Inhibition of rat hepatocyte in multiplication by serum factors. Virchows Arch [Cell Pathol] 18:273–280Google Scholar
  49. Nadal C (1967) Poliploïdie hépatique du rat. III. Etude des nucléoles des cellules hépatiques du rat normal. Exp Cell Res 48:507–517CrossRefGoogle Scholar
  50. Nadal C, Zajdela F (1967) Polyploidie hépatique du rat. IV. Variations expérimentales du volume nucléolaire des hépatocytes et leurs mécanismes de régulation. Exp Cell Res 48:518–528PubMedCrossRefGoogle Scholar
  51. Ord MG, Stocken LLA (1980) Nuclear-protein changes in the cell cycle: interphase studies with regenerating rat liver. Biochem Soc Trans 8:759–766PubMedGoogle Scholar
  52. Prescott DM (1969) Sequential events of cell life cycle. In: Exploitable molecular mechanisms and neoplasia. Williams and Wilkins, Baltimore, pp 359–370Google Scholar
  53. Reynolds ES (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17:208–212PubMedCrossRefGoogle Scholar
  54. Rohr HP, Strebel J, Bianchi L (1970) Ultrastructural morphometric study on rat liver cells in the early regenerative phase after partial hepatectomy. Beitr Pathol 141:52–74PubMedGoogle Scholar
  55. Serratosa J (1985) Caracterización estructural y ultraestructural de la regeneratión hepática en la rata. Estudio morfométrico. Doct Thesis. Universidad de Barcelona, pp 200–206Google Scholar
  56. Steiner JW, Perz ZM, Taichman LB (1966) Cell population dynamics in liver: Review of quantitative morphological techniques applied to study of physiological and pathological growth. Exp Mol Pathol 5:146–181PubMedCrossRefGoogle Scholar
  57. Stenger RJ, Confer DB (1966) Hepatocellular ultrastructure during liver regeneration after subtotal hepatectomy. Exp Mol Pathol 5:455–457PubMedCrossRefGoogle Scholar
  58. Stowell RE (1948) Nucleic acids and cytologic changes in regenerating rat liver. Arch Pathol (Chicago) 46:164–178Google Scholar
  59. Takatsuka Y, Kohno M, Higashi K, Hirano H, Sakamoto Y (1976) Redistribution of chromatin ribosomal cistrons durin liver regeneration. Exp Cell Res 103:191–199PubMedCrossRefGoogle Scholar
  60. Tsukada K, Lieberman I (1964) Metabolism of nuclear ribonucleic acid after partial hepatectomy. J Biol Chem 239:1564–1568PubMedGoogle Scholar
  61. Tubo RA, Berezney R (1987) Pre-replicative association of multiple replicative enzyme activities with the nuclear matrix during liver regeneration. J Biol Chem 262:1148–1154PubMedGoogle Scholar
  62. Tucker RW, Pardee AB (1982) Prymary cilia and their role in the regulation of DNA replication and mitosis. In: Nicolini MA (ed) Cell growth. Plenum Press, New York and London, pp 365–376Google Scholar
  63. Virolainen M (1967) Humoral control mechanism in liver regeneration. Helsinki. Painovalmiste, pp 1–105Google Scholar
  64. Weibel ER, Paumgartner D (1978) Integrated stereologycal and biochemical studies on hepatocytic membranes. II. Correction of section thickness effect on volumen surface density stimates. J Cell Biol 584–597Google Scholar
  65. Weibel ER, Kistler GS, Scherle WF (1966) Practical Stereological methods for morphometric cytology. J Cell Biol 30:23–38PubMedCrossRefGoogle Scholar
  66. Weibel ER, Staubli W, Gnagi HR, Hess FA (1969) Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, stereologic methods and normal morphometric data for rat liver. J Cell Biol 42:69–91CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Joan Serratosa
    • 1
  • Jordi Domingo
    • 2
  • Carles Enrich
    • 2
  • Oriol Bachs
    • 2
  1. 1.Departamento de Farmacología y ToxicologiaCentro de Investigatión y Desarrollo del C.S.I.C.Barcelona
  2. 2.Departamento de Biologia Celular y Anatomia Patológica, Facultad de MedicinaUniversidad de BarcelonaBarcelonaSpain

Personalised recommendations