Abstract
Differentiation of primitive erythroid cells derived from the yolk sac of the chick embryo is accompanied by changes in the morphology of and in the physicochemical properties of the nucleus. Microfluorimetry of individual nuclei stained with acridine orange was performed on thermally denatured cells. Measurements were made at 530 nm (green fluorescence) and 590 nm (redfluorescence). The ratio of these two measurements was used to monitor the susceptibility of chromatin to thermal denaturation. Differences were found (a) between mature erythrocytes and dividing erythroblasts, and (b) between dividing erythroblasts from successive cell generations of the erythroid series. There were differential characteristics of AO binding during thermal denaturation as signified by F530 and F590 measurements. The temperature at which the increase of the “α ratio” (F590/F530) was 50% of its maximum was approximately 70° C for erythroblasts from the fifth generation (day 4), 80–85° C for the sixth generation (day 5), and 85–90° C for the nondividing erythrocytes (day 8). Interpretation of these differences may be complicated by changes in the sensitivity of nuclear proteins to the interactive effects of 0.15 M NaCl and thermal denaturation.
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Appels, R., Wells, J. R. E., William, A. F.: Characterization of DNA-Bound histone in the cells of the avian erythropoietic series. J. Cell Sci. 10, 47–59 (1972).
Auer, G., Zetterberg, A., Killander, D.: Changes in binding between DNA and arginine residues in histone induced by cell crowding. Exp. Cell Res. 62, 32–38 (1970).
Campbell, G. LeM., Weintraub, H., Mayall, B. H., Holtzer, H.: Primitive erythropoiesis in early chick embryogenesis. II. Correlation between hemoglobin synthesis and the mitotic history. J. Cell Biol. 50, 669–681 (1971).
Dreskin, S. C., Mayall, B. H.: Thermal denaturation of chromatin in fixed cells: Changes in staining with Gallocyanin-ehrome alum (GCA) and in digestibility with a single strand-specific DNase (SSDNase). J. Histochem. Cytochem. 19, 718 (Abstract) (1971).
Grogan, D. E., Desjardins, R., Busch, H.: Nucleolar proteins of rat liver and Walker tumor. Cancer Res. 26, 775–779 (1966).
Kernell, A. M., Bolund, L., Ringertz, N. R.: Chromatin changes during erythropoiesis. Exp. Cell Res. 65, 1–6 (1971).
Lucas, A. M., Jamroz, C.: Atlas of avian hematology, p. 25. Washington, D. C.: U. S. Department of Agriculture, 1961.
Richards, B. M., Pardon, J. F.: The molecular structure of nucleohistone (DNH). Exp. Cell Res. 62, 185–196 (1970).
Rigler, R.: Microfluorometric characterization of intracellular nucleic acids and nueleoproteins by acridine orange. Acta physiol. scand. 67, Suppl. 267, 1–122 (1970).
Rigler, R., Killander, D., Bolund, L., Ringertz, N. R.: Cytoehemical characterization of deoxyribonucleoprotein in individual cell nuclei. Exp. Cell Res. 55, 215–224 (1969).
Ringertz, N. R.: Cytochemical properties of nuclearproteins and deoxyribunucleoproteincomplexes in relation to nuclear function. In: Handbook of molecular cytology (A. Lima-de-Faria, ed.), pp. 657–684. Amsterdam: North Holland Publishing Co., 1969.
Ringertz, N. R., Gledhill, B. L., Darzynkiewicz, Z.: Changes in deoxyribonucleoprotein during sperminogenesis in the bull: Sensitivity of DNA to heat denaturation. Exp. Cell. Res. 62, 204–218 (1970).
Seligy, V. L., Neelin, J. M.: Transcription properties of stepwise acid-extracted chicken erythrocyte chromatin. Biochim. biophys. Acta (Amst.) 213, 380–390 (1970).
Smart, J. E., Bonner, J.: Studies on the role of histones in relation to the template activity and precipitability of chromatin at physiological ionic strengths. J. molec. Biol. 58, 675–684 (1971).
Steele, W. J., Busch, H.: Studies on acidic nuclear proteins of the Walker tumor and liver. Cancer Res. 23, 1153–1163 (1963).
Weintraub, H., Campbell, G. LeM., Holtzer, H.: Primitive erythropoiesis in early chick embryogeneisis. I. Cell cycle kinetics and the control of cell division. J. Cell Biol. 50, 652–668 (1971).
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Campbell, G.L., Gledhill, B.L. Chromatin of primitive erythroid cells from the chick embryo. Chromosoma 41, 385–394 (1973). https://doi.org/10.1007/BF00396496
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DOI: https://doi.org/10.1007/BF00396496