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Étude en microscopie électronique et par cytophotométrie à balayage de la structure et de la distribution de la chromatine dans les noyaux des cellules cartilagineuses deTriturus cristatus âgés au cours de la régénération

Electron microscopic and scanning cytophotometric study of chromatin structure and distribution in nuclei of cartilaginous cells of agedTriturus cristatus during regeneration

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Summary

Cartilaginous cells of aged newts (Triturus cristatus) were studied during hind limb regeneration. The electron microscope was used to study the structure and distribution of chromatin in the cell nuclei, while the DNA content of the chromatin was measured by means of a scanning cytophotometer.

Changes in the ultrastructure of the cytoplasm during regeneration were also studied.

It was observed that the structure and distribution of chromatin in the activated cell is greatly modified. In the non-activated cell of the aged newt, the chromatin is found highly condensed and distributed peripherally close to the nuclear membrane. In contrast, in the activated cells, the chromatin is much less condensed and is distributed throughout the nucleus. Moreover, cytoplasmic vacuoles, found only in the non-activated aged cells, disappear and an increase in the mitochondria and rough endoplasmic reticulum is also observed.

Changes in the nuclear structure are observed prior to the cytoplasmic modifications.

It is interesting to note that the process of activation induces structural changes in the aged cells which make these cells appear to be structurally identical to the young cells. This process of rejuvenation takes 3–5 days in the newt.

We suggest that these structural changes of the chromatin and cytoplasm in the aged cells are necessary to increase the metabolic activity which precedes cell division. It may also explain why regeneration takes a longer time in the aged animals than in the young ones.

Résumé

Les cellules cartilagineuses des membres postérieurs deTriturus cristatus en régénération après amputation, ont été étudiées en microscopie électronique et par cytophotométrie à balayage. Nous nous sommes intéressés à la structure et à la distribution de la chromatine mais aussi à différents organites cytoplasmiques. Dans l'étude de cytophotométrie à balayage, la chromatine a été considérée à travers son constituant majeur, l'ADN, coloré par la réaction de Feulgen. Au cours de la régénération du membre, l'hétérochromatine initialement condensée, essentiellement accolée à la membrane nucléaire se décondense. Les vacuoles du cytoplasme, caractéristiques des animaux âgés par rapport aux animaux jeunes, disparaissent, les mitochondries et le reticulum endoplasmique rugueux deviennent plus abondants. Les caractéristiques nucléaires de l'activation cellulaire apparaissent précocement, précédent les modifications cytoplasmiques et conduisent à des cellules en tous points identiques aux cellules d'animaux jeunes en dehors de tout processus régénératif. Cette phase d'euchromatisation et de restructuration cytoplasmique est peut-être nécessaire à l'accroissement d'activité métabolique et à la division cellulaire qui suivent. Son déroulement peut expliquer tout au moins le ralentissement de la régénération observé chez les animaux âgés par rapport aux animaux jeunes.

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Bibliographie

  • Anton, H.J.: Autoradiographische Untersuchungen über den Eiweißstoffwechsel bei der Extremitä-tenregeneration der Urodelen. Wilhelm Roux' Arch. Entwickl.-Mech. Org.161, 49–88 (1968)

    Google Scholar 

  • Arnold, E.A., Yawn, D.H., Brown, D.G., Wyllie, R.C., Coffey, D.S.: Structural alteration in isolated rat liver nuclei after removal of template restriction by polyanions. J. Cell Biol.53, 737–757 (1972)

    PubMed  Google Scholar 

  • Augenlicht, L.H., Baserga, R.: Changes in the G0 state of WI-38 fibroblasts at different times after confluence. Exp. Cell Res.89, 255–262 (1974)

    PubMed  Google Scholar 

  • Berger, N.A., Skinner, Sr.A.M.: Characterization of lymphocyte transformation induced by zinc ions. J. Cell Biol.61, 45–55 (1974)

    Google Scholar 

  • Bolund, L., Ringertz, N.R., Harris, H.: Changes in the cytochemical properties of erythrocyte nuclei reactivated by cell fusion. J. Cell Sci.4, 71–87 (1969)

    PubMed  Google Scholar 

  • Brasch, K., Seligy, V.L., Setterfield, G.: Effects of low salt, concentration on structural organization and template activity of chromatin in chicken erythrocyte nuclei. Exp. Cell. Res.65, 61–72 (1971)

    PubMed  Google Scholar 

  • Christov, K., Kiefer, G., Kiefer, R., Sandritter, W.: Changes in the nuclear structure during thyroid carcinogenesis in rats. An image analysis study. Beitr. Pathol.152, 19–36 (1974)

    PubMed  Google Scholar 

  • Darda, S., Anton, H.J.: Quantitative Untersuchungen über die Proteinsyntheseaktivität von Epidermiszellen während der Extremitätenregeneration der Urodelen. Experientia25, 1321–1322 (1969)

    PubMed  Google Scholar 

  • Darzinkiewicz, Z., Bolund, L., Ringertz, N.R.: Nucleoprotein changes and initiation of RNA synthesis in PHA stimulated lymphocytes. Exp. Cell Res.56, 418–424 (1969)

    PubMed  Google Scholar 

  • Deitch A.D.: Cytophotometry of nucleic acids. In: Introduction to quantitative cytochemistry (G.L. Wied, ed.), pp. 327–354 New York and London: Academic Press 1966

    Google Scholar 

  • Desselle, J.C.: Cytophotométrie des acides nucléiques dans les cellules musculaires de membres en régénération, de membres irradiés aux rayons X, de membres irradiés réactivés par implant de cartilage chezTriturus cristatus. Acta Embryol. exp.3, 207–235 (1974)

    Google Scholar 

  • Desselle, J.C.: Analyse cytochimique des histones des cellules musculaires des membres deTriturus cristatus en régénération normale, irradiés par les rayons X et au cours de la restauration de la régénération par des implants. Compt. Rend., Paris282, 301–304 (1976)

    Google Scholar 

  • Dupuy-Coin, A.M., Ege, T., Bouteille, M., Ringertz, N.R.: Ultrastructure of chick erythrocyte nuclei undergoing reactivation in heterokaryons and enucleated cells. Exp. Cell Res.101, 355–369 (1976)

    PubMed  Google Scholar 

  • Elves, M.W., Gough, J., Chapman, J.A., Israels, M.C.G.: Electron microscopy studies of lymphocytes. Transformation under the influence of phytohaemagglutinin. Lancet1, 306–308 (1964)

    Google Scholar 

  • Fermanian, J., Salmon, D.: Mathématiques P.C.E.M. Statistique. In: Synthèse,31, 112 p. Paris: Armand Colin, 1974

    Google Scholar 

  • Firket, H.: Evolution ultrastructurale de lymphocytes humains cultivés en présence de phytohémagglutinine (PHA). J. Microsc.5, 48a (1966)

    Google Scholar 

  • Firket, H.: L'évolution de l'ultrastructure du lymphocyte humain en culture sous l'influence de la phytohémagglutinine. Comparaison avec le lymphocyte en culture mixte. Nouv. Rev. Fr. Hematol.9, 159–176 (1969)

    PubMed  Google Scholar 

  • Gabie, V., Andrew, A.: A cytochemical study of histone changes during lens regeneration in the newt.Triturus (Diemyctylus) viridescens. Acta Embryol. Morphol. Exp.10, 31–43 (1967)

    PubMed  Google Scholar 

  • Gurdon, J.B.: Changes in somatic cell nuclei inserted into growing and maturing amphibian oocytes. J. Embryol. Exp. Morphol.20, 401–414 (1968)

    PubMed  Google Scholar 

  • Gurdon, J.B., Weir, R.S.: Cytoplasmic proteins and the control of nuclear activity in early amphibian development. Biochem. J.114, 52P-53P (1969)

    PubMed  Google Scholar 

  • Gurdon, J.B., Woodland, H.R.: The cytoplasmic control of nuclear activity in animal development. Biol. Rev.43, 233–267 (1968)

    PubMed  Google Scholar 

  • Harris, H.: The reactivation of the red cell. J. Cell Sci.2, 23–32 (1967)

    PubMed  Google Scholar 

  • Hay, E.D.: The fine structure of blastema cells and differentiating cartilage cells in regenerating limbs ofAmblystoma larvae. J. Biophys. Biochem. Cytol.4, 583–591 (1958)

    PubMed  Google Scholar 

  • Hay, E.D.: Electron microscopic observations of muscle dedifferentiation in regeneratingAmblystoma limbs. Dev. Biol.1, 555–585 (1959)

    Google Scholar 

  • Hay, E.D.: Cytological studies of dedifferentiation and differentiation in regenerating amphibian limbs. In: Regeneration (D. Rudnick, ed.), pp. 177–210 New York: Ronald Press Company, 1962

    Google Scholar 

  • Inman, D.R., Cooper, E.H.: Electron microscopy of human lymphocytes stimulated by phytohaemagglutinin. J. Cell Biol.19, 441–445 (1963)

    PubMed  Google Scholar 

  • Jeanny, J.C.: Etude cytophotométrique des acides nucléiques et des histones des cellules cartilagineuses activées au cours de la régénération du membre deDesmognathus fuscus (Amphibien, Urodèle, Pléthodontidé). Ann. Embryol. Morph.,6, 25–41 (1973)

    Google Scholar 

  • Jeanny, J.C.: Modifications des propriétés cytophotométriques des ADN et des histones nucléaires au cours de la sénescence des tritons (Triturus vulgaris etTriturus cristatus). Exp. Cell Res.102, 394–404 (1976)

    PubMed  Google Scholar 

  • Kiefer, R., Kiefer, G., Salm, R., Rossner, R., Sandritter, W.: A method for the quantitative evaluation of eu- and heterochromatin in interphase nuclei using cytophotometry and pattern analysis. Beitr. Pathol.150, 163–173 (1973)

    PubMed  Google Scholar 

  • Kiefer, G., Kiefer, R., Moore, G.W., Salm, R., Sandritter, W.: Nuclear images of cells in different functional states. J. Histochem. Cytochem.22 569–576 (1974)

    PubMed  Google Scholar 

  • Kiefer, G., Sandritter, W.: DNA and the cell cycle. Beitr. Pathol.158, 332–362 (1976)

    PubMed  Google Scholar 

  • Luft, J.H.: Improvements in epoxy resin embedding methods. J. Biophys. Biochem. Cytol.9, 409–414 (1961)

    PubMed  Google Scholar 

  • Miller, G., Berlowitz, L., Regelson, W.: Chromatin and histones in mealy bug cell explants: activation and decondensation of facultative heterochromatin by a synthetic polyanion. Chromosoma32, 251–261 (1971)

    PubMed  Google Scholar 

  • Morzlock, F.V., Stocum, D.L.: Patterns of RNA synthesis in regenerating limbs of the adult newt,Triturus viridescens. Dev. Biol.24, 106–118 (1971)

    PubMed  Google Scholar 

  • 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 (1963)

    Article  PubMed  Google Scholar 

  • Rowinski, J., Pienkowski, M., Abramczuk, J.: Area representation of optical density of chromatin in resting and stimulated lymphocytes as measured by means of quantimet. Histochemie32, 75–80 (1972)

    PubMed  Google Scholar 

  • Rowinski, J. Sawicki, W., Swenson, R., Koprowski, H.: Changes in chromatin morphology after infection of mouse embryo fibroblasts with polyoma virus, detected by image analysis. Acta Cytol.19, 136–141 (1975)

    PubMed  Google Scholar 

  • Sabatini, D.D., Bensch, K., Barrnett, R.J.: Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol.17, 19–58 (1963)

    PubMed  Google Scholar 

  • Salpeter, M.M., Singer, M.: The fine structure of mesenchymatous cells in the regenerating forelimb of the adult newtTriturus. Dev. Biol.2, 516–534 (1960)

    PubMed  Google Scholar 

  • Salpeter, M.M., Singer, M.: The fine structure of mesenchymatous cells in regenerating limbs of larval and adultTriturus. In: Electron microscopy. Fifth international congress for electron microscopy. (S.S. Breese (ed.),2, pp. 00–12 New York and London: Academic Press, 1962

    Google Scholar 

  • Sandritter, W., Pilny, J., Novakova, V., Kiefer, G.: Zur Problematik der Gewebspräparation für cytophotometrische Messungen. Histochemie7, 1–7 (1966)

    PubMed  Google Scholar 

  • Sandritter, W., Kiefer, G.: Objectivization of chromatin patterns using the fast-scanning stage of the UMSP-I. In: Automated cell identification and cell sorting (G.L. Wied and G.F. Bahr, eds.), pp. 177–185. New York and London, Academic Press, 1970

    Google Scholar 

  • Sandritter, W., Kiefer, G., Kiefer, R., Salm, R., Moore, G.W., Grimm, H.: DNA in heterochromatin cytophotometric pattern recognition image analysis among cell nuclei in duct epithelium and in carcinoma of the human breast. Beitr. Pathol.151, 87–96 (1974)

    PubMed  Google Scholar 

  • Sawicki, W., Rowinski, J., Abramczuk, J.: Image analysis of chromatin in cells of preimplantation mouse embryos. J. Cell Biol.63, 227–233 (1974a)

    PubMed  Google Scholar 

  • Sawicki, W., Rowinski, J., Swenson, R.: Change of chromatin morphology during the cell cycle detected by means of automated image analysis. J. Cell Physiol.84, 423–428 (1974b)

    PubMed  Google Scholar 

  • Sprenger, E., Moore, G.W., Naujoks, H., Schlüter, G., Sandritter, W.: DNA content and chromatin pattern analysis on cervical carcinoma in situ. Acta Cytol.17, 27–31 (1973)

    PubMed  Google Scholar 

  • Watson, M.L.: Staining of tissue sections for electron microscopy with heavy metals. J. Biophys. Biochem. Cytol.4, 475–478 (1958)

    PubMed  Google Scholar 

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Jeanny, J.C., Gontcharoff, M. Étude en microscopie électronique et par cytophotométrie à balayage de la structure et de la distribution de la chromatine dans les noyaux des cellules cartilagineuses deTriturus cristatus âgés au cours de la régénération. Wilhelm Roux' Archiv 184, 195–211 (1978). https://doi.org/10.1007/BF00848254

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