, Volume 55, Issue 1, pp 63–74 | Cite as

Microdensitometric and autoradiographic comparison of the DNA contents of foetal and adult rat liver nuclei

  • K. S. Bedi
  • D. J. Goldstein


Glare-corrected, scanning Feulgen microdensitometry and 3H-thymidine autoradiography were applied to squash preparations of rat 18-day foetal and maternal liver cells, and to smears of maternal blood. No significant differences were found between the mean Feulgen-DNA contents of autoradiographically unlabelled diploid foetal and maternal hepatocytes. The Feulgen-DNA contents of other unlabelled foetal and maternal hepatocytes were also as predicted by the DNA-constancy hypothesis, i.e. were twice or four times that of diploid cells. Small (less than about 4%) but statistically significant discrepancies in the mean Feulgen-DNA contents of foetal haemopoietic cells and adult leucocytes were attributable to uncorrected residual distribution and chromatic errors in the microdensitometry.

None of the 371 maternal nuclei measured had Feulgen-DNA contents substantially (i.e. more than ±10%) different from a modal value. About 12% of these nuclei were classified as labelled. Evidence was found suggesting a significantly non-random distribution of ‘background” grains in the autoradiographs, which would materially affect the proportion of cells incorrectly classified. After taking this factor into account there seems no reason to suppose that the apparently labelled adult nuclei were in fact synthesising DNA.

Of 376 foetal cells measured, 107 had inter-modal Feulgen-DNA contents. Eleven of these were classified as unlabelled. All the inter-modal cells were however probably in the S-phase of the cell cycle, statistical variation in autoradiographic grain distribution accounting for those appearing to be unlabelled.

Our results are consistent with the DNA-constancy hypothesis, and are at variance with previous claims for the existence of ‘metabolic’ DNA in adult and/or foetal rat hepatocytes.


Maternal Blood Diploid Cell Foetal Cell Residual Distribution Previous Claim 
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  1. Arold, R., Sandritter, W.: Zytophotmetrische Bestimmungen des DNS-Gehaltes von Zellkernen des Nebennierenmarkes, der Nebennierenrinde und der Schilddrüse unter verschiedenen experimentellen Bedingungen, Histochemie 10, 88–97 (1967)Google Scholar
  2. Bedi, K.S., Goldstein, D.J.: Apparent anomalies in nuclear Feulgen-DNA contents. Role of systematic microdensitometric errors. J. Cell Biol. 71, 68–88 (1976)Google Scholar
  3. Benedetti, P.A., Viola-Magni, M.P.: A scanning integrating histophotometer. J. Sci Instrum. 43, 141–143 (1966)Google Scholar
  4. Bibbiani, C., Tongiani, R., Viola-Magni, M.P.: I. Quantitative determination of the amount of DNA per nucleus by interference microscopy. J. Cell Biol. 42, 444–451 (1969)Google Scholar
  5. Bibbiani, C., Tongiani, R., Viola-Magni, M.P.: Changes of DNA content per nucleus in hepatocytes of rat during the first days of Postnatal life. Histochemie 35, 189–198 (1973)Google Scholar
  6. Böhm, N., Sprenger, E., Schlüter, G., Sandritter, W.: Proportionalitätsfehler bei der Feulgen Hydrolyse. Histochemie 15, 194–203 (1968)Google Scholar
  7. Boivin, A., Vendrely, R., Vendrely, C.: L'acide désoxyribonucléique due noyau cellulaire dépositaire des caractères héréditaires; arguments d'ordre analytique. C.r. Hebd. Séanc. Acad. Sci. Paris. 226, 1061–1063 (1948)Google Scholar
  8. Cleaver, J.E.: Thymidine metabolism and Cell Kinetics (ed. A. Neuberger and E.L. Tatum). Amsterdam: North Holland Publ. 1964Google Scholar
  9. Cohn, N.S., van Duijn, P.: Constancy of DNA content in adrenal medulla nuclei of cold-treated rats. J. Cell Biol. 33, 349–354 (1967)Google Scholar
  10. Dempster, W.J. and M.A. Williams. (1963). Cellular infiltration in homotransplanted kidneys. Brit. Med. J. 1963/I, 18–23Google Scholar
  11. Dendy, P.P., Cleaver, J.E.: An investigation of (a) variation in rate of DNA-synthesis during S-phase in mouse L-cells; (b) effect of UV radiation on rate of DNA-synthesis. Intern J. Radiat. Biol. 8, 301–315 (1964)Google Scholar
  12. Fukuda, M., Sibatani, A.: Biochemical studies on the number and the composition of liver cells in postnatal growth of the rat. J. Biochem. (Tokyo) 40, 95–110 (1953a)Google Scholar
  13. Fukuda, M., Sibatani, A.: Relation between the body wt and the average DNA content of liver nuclei in postnatal growth of the rat. Exp. Cell Res. 4, 236–238 (1953b)Google Scholar
  14. Gall, J.G., Johnson, W.W.: Is there “Metabolic” DNA in the mouse seminal vesicle? J. Biophys. Biochem Cytol. 7, 657–664 (1960)Google Scholar
  15. Goldstein, D.J.: Aspects of scanning microdensitometry. I. stray light (glare). J. Microsc. 92, 1–16 (1970)Google Scholar
  16. Goldstein, D.J.: Aspects of scanning microdensitometry. II. Spot size, focus and resolution. J. Microsc. 93, 15–42 (1971)Google Scholar
  17. Goldstein, D.J.: Aspects of scanning microdensitometry. III. The monochromator system. J. Microsc. 105, 33–56 (1975)Google Scholar
  18. Harris, G.: DNA synthesis and the production of antibodies by lymphoid tissues. Differentiation 1, 301–318 (1973a)Google Scholar
  19. Harris, G.: The immune response of spleen explants from primed rabbits to sheep red cells. Immunology 24, 343–363 (1973b)Google Scholar
  20. Harris, G.: Autoradiographic studies of DNA synthesis in lymphoid tissues. J. Microsc. 106, 173–184 (1976)Google Scholar
  21. Harris, G., Pelc, S.R.: Incorporation of 3H thymidine into the spleens of intact mice during the immune response to sheep erythrocytes. Immunology 19, 865 (1970)Google Scholar
  22. James, J.: Constancy of nuclear DNA and accurary of cytophotometric measurements. Cytogenetics 4, 19–27 (1965)Google Scholar
  23. James, J.: Feulgen-DNA changes in rat liver cell nuclei during the early phase of ischaemic necrosis. Histochemie 13, 312–322 (1968)Google Scholar
  24. Mirsky, A.E., Ris, H.: Variable and constant components of chromosomes. Nature (London) 163, 666–667 (1949)Google Scholar
  25. Naora, H.: Schwarzchild-Villiger effect in microspectrophotometry. Science, 115, 248–249 (1952)Google Scholar
  26. Naora, H.: Microspectrophotometry of cell nucleus stained by Feulgen reaction. Exp. Cell Res. 8, 259–278 (1955)Google Scholar
  27. Naora, H.: Microspectrophotometry of cell nuclei stained with Feulgen reaction. iii) The DNA content of individual nuclei of rat tissues in the postnatal growth. Exp. Cell Res. 12, 1–14 (1957)Google Scholar
  28. Pelc, S.R.: Labelling of DNA and cell division in so called non-dividing tissues. J. Cell Biol. 22, 21–28 (1964)Google Scholar
  29. Pelc, S.R.: Metabolic DNA in ciliated protozoa, salivary gland chromosomes, and mammalian cells. Int. Rev. Cytol. 32, 327–355 (1972)Google Scholar
  30. Pelc, S.R., Gahan, P.B.: Incorporation of labelled thymidine in the seminal vesicle of the mouse. Nature (London) 183, 335–336 (1959)Google Scholar
  31. Pelc, S.R., Viola-Magni, M.P.: III. Decrease of labelled DNA in cells of the adrenal medulla after intermittent exposure to cold. J. Cell Biol. 42, 460–468 (1969)Google Scholar
  32. Potter, C.G.: Microspectrophotometry and autoradiography in the study of DNA values of cells in tissue culture. Exp. Cell Res. 61, 141–146 (1970)Google Scholar
  33. Roels, H.: “Metabolic” DNA: A cytochemical study. Int. Rev. Cytol. 19, 1–34 (1966)Google Scholar
  34. Rogers, A.W.: Techniques of autoradiography. Amsterdam and New York: Elsevier Publishing Co. 1966Google Scholar
  35. Tongiani, R.: An integrating apparatus for microinterferometry. Acta Histochem. 33, 13–29 (1969)Google Scholar
  36. Tongiani, R., Viola-Magni, M.P.: II. Differences in adrenal medulla nuclear DNA content among rats of different strains following intermittent exposure to cold. J. Cell Biol. 42, 452–459 (1969)Google Scholar
  37. Viola-Magni, M.P.: A radioautographic study with H3-thymidine on adrenal medulla nuclei of rats intermittently exposed to cold. J. Cell Biol. 28, 9–19 (1966)Google Scholar
  38. Viola-Magni, M.P.: Synthesis and turnover of DNA in hepatocytes of neonatal rats. J. Microsc. 96, 191–203 (1972)Google Scholar

Copyright information

© Springer-Verlag 1978

Authors and Affiliations

  • K. S. Bedi
    • 1
  • D. J. Goldstein
    • 1
  1. 1.Department of Human Biology and AnatomyUniversity of SheffieldSheffieldEngland

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