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Chromatin behaviour under influence of puromycin and 6-DMAP at different stages of mouse oocyte maturation

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Abstract

Preovulatory mouse oocytes were cultured in vitro up to each subsequent stages of maturation: germinal vesicle (GV), germinal vesicle breakdown (GVBD), groups of not yet individualized bivalents, circular bivalents, late prometaphase I, metaphase I, anaphase I and telophase I. The stages were identified in living oocytes by fluorescence microscopy using Hoechst 33342 as a specific vital dye. Oocytes from each stage of development developed in vitro and ovulated metaphase II oocytes were subsequently cultured in the presence of puromycin or 6-dimethylaminopurine (6-DMAP), an inhibitor of protein phosphorylation. The effects on chromatin of these drugs were studied during and at the end of culture by fluorescence and electron microscopy. We found that puromycin and 6-DMAP stop meiosis when applied at all stages of oocyte maturation, except for metaphase II. Oocytes at this stage are activated by puromycin. Reaction of the oocytes to the two drugs is different at GV and at metaphase II. All of the other stages react to the drugs by chromatin compaction, which can be followed by chromatin decondensation to form a nucleus. Our results suggest that late prophase chromatin condensation, bivalent individualization and retention of their individuality, as well as individualization of monovalents from telophase and retention of their individuality at metaphase II, are dependent on protein phosphorylation. The events occurring between metaphase I and telophase I are independent of protein synthesis and phosphorylation. The events occurring between metaphase II and formation of the nucleus are independent of protein synthesis.

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Authors and Affiliations

  1. Unité de Biologie de la Fécondation, Station de Physiologie Animale, I.N.R.A., F-78352, Jouy-en-Josas Cédex, France

    Maria S. Szöllösi & Daniel Szöllösi

  2. Unité en Développement concerté I.N.S.E.R.M.-I.N.R.A. U.310, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, F-75005, Paris, France

    Pascale Debey & Dominique Vautier

  3. Laboratoire de Physiologie de la Reproduction, Groupe Stéroïdes, I.N.R.A.-C.N.R.S., UA 555, Université Pierre et Marie Curie, 4 place Jussieu, F-75252, Paris Cédex 05, France

    Hélène Rime

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  1. Maria S. Szöllösi
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  2. Pascale Debey
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  3. Daniel Szöllösi
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  4. Hélène Rime
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  5. Dominique Vautier
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by U. Scheer

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Szöllösi, M.S., Debey, P., Szöllösi, D. et al. Chromatin behaviour under influence of puromycin and 6-DMAP at different stages of mouse oocyte maturation. Chromosoma 100, 339–354 (1991). https://doi.org/10.1007/BF00360533

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  • DOI: https://doi.org/10.1007/BF00360533

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