Summary
In the absence of a suitable energy source, mouse oocytes cultured in vitro resume, but fail to complete, meiotic maturation. However, little is known about the underlying mechanisms leading to this meiotic failure. We utilized pyruvate-deficient medium to test for the role of pyruvate throughout the meiotic maturation process. Germinal vesicle-stage (GV) oocytes underwent germinal vesicle breakdown (GVBD), but failed to form a polar body when cultured continuously in pyruvate-free medium. However, when GV oocytes were preincubated for 4 h in pyruvate-free medium containing dibutyryl cyclic adenosine monophosphate (dbcAMP) and then cultured in pyruvate-free medium, GVBD was markedly inhibited. Preincubation of GV oocytes in dbcAMP and cycloheximide, followed by culture in cycloheximide only, also inhibited GVBD. A longer preincubation period was required in the cycloheximide-dbcAMP case (12 h) than in pyruvate-free-dbcAMP medium situation (4 h). Strikingly, reassembly of the nuclear membrane without polar body formation was observed following GVBD in oocytes continuously cultured in pyruvate-free medium. The reassembled nuclear membrane increased in size with continued culture, and it surrounded partially-decondensed chromatin. Nuclear membrane reassembly also occurred in oocytes which had undergone GVBD during continuous culture in medium containing only cycloheximide. Reformation of nuclear membranes after GVBD was confirmed by electron-microscopic analyses of oocytes cultured in pyruvate-free medium or in the presence of cycloheximide. We conclude that both pyruvate and protein synthesis are required for nuclear membrane disassembly, whereas lack of pyruvate or protein synthesis is associated with interruption of the metaphase state and reassembly of the nuclear membrane. The evidence suggests that assembly and maintenance of an intact nucleus and its disintegration are all amenable to regulation by pyruvate, possibly via mechanism(s) involving protein synthesis.
Similar content being viewed by others
References
Biggers JD, Whittingham DG, Donahue RP (1967) The pattern of energy metabolism in the mouse and zygote. Proc Natl Acad Sci USA 58:560–567
Blerkom J van (1981) The structural relationship and post-translational modification of stage-specific proteins synthesized during early preimplantation development in the mouse. Proc Natl Acad Sci USA 78:7629–7633
Cascio SM, Wassarman PM (1982) Program of early development in the mammal: Posttranscriptional control of a class of proteins synthesized by mouse oocytes and early embryos. Dev Biol 89:397–408
Cho WK, Stern S, Biggers JD (1974) Inhibitory effect of dibutyryl cAMP on mouse oocyte maturation in vitro. J Exp Zool 187:383–386
Clarke HJ, Masui Y (1983) The induction of reversible and irreversible chromosome decondensation by protein synthesis inhibition during meiotic maturation of mouse oocytes. Dev Biol 97:291–301
Clarke HJ, Rossant J, Masui Y (1988) Suppression of chromosome condensation during meiotic maturation induces parthenogenetic development of mouse oocytes. Development 104:97–103
Donahue RP (1968) Maturation of the mouse oocyte in vitro. I. Sequence and timing of nuclear progression. J Exp Zool 169:237–250
Donahue RP, Stern S (1968) Follicular cell support of oocyte maturation: production of pyruvate in vitro. J Reprod Fert 17:395–398
Ekholm C, Magnusson C (1979) Rat oocyte maturation: effects of protein synthesis inhibitors. Biol Reprod 21:1287–1293
Golbus MS, Stein MP (1976) Qualitative patterns of protein synthesis in the mouse oocyte. J Exp Zool 198:337–342
Hashimoto N, Kishimoto T (1988) Regulation of meiotic metaphase by a cytoplasmic maturation-promoting factor during mouse oocyte maturation. Dev Biol 126:242–252
Kim H, Schuetz AW (1991) Regulation of parthenogenetic activation of metaphase II mouse oocytes in vitro by pyruvate. J Exp Zool 251:375–385
Krishan A (1975) Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol 66:188–193
Kubelka M, Motlik J, Fulka J Jr, Prochazka R, Rimkevicova Z, Fulka J (1988) Time sequence of germinal vesicle breakdown in pig oocytes after cycloheximide and p-aminobenzamidine block. Gamete Res 19:423–431
Masui Y (1985) Meiotic arrest in animal oocytes. In: Metz CB, Monroy A (eds) Biology of fertilization. Academic Press, New York, pp 189–219
Newport J, Spann T (1987) Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes. Cell 48:219–230
Rime H, Neant I, Guerrier P, Ozon R (1989) 6-Dimethyl-aminopurine (6-DMAP), a reversible inhibitor of the transition to metaphase during the first meiotic cell division of the mouse oocyte. Dev Biol 133:169–179
Schuetz AW (1985) Local control mechanisms during oogenesis and folliculogenesis. In: Browder LW (ed) Developmental Biology. Plenum Press, New York, pp 3–83
Schultz RM, Wassarman PM (1977a) Biochemical studies of mammalian oogenesis: Protein synthesis during oocyte growth and meiotic maturation. J Cell Sci 24:167–194
Schultz RM, Wassarman PM (1977b) Specific changes in the pattern of protein synthesis during meiotic maturation of mammalian oocytes in vitro. Proc Natl Acad Sci USA 74:538–541
Siracusa G, Whittingham DG, Molinaro M, Vivarelli E (1978) Parthenogenetic activation of mouse oocytes induced by inhibitors of protein synthesis. J Embryol Exp Morphol 43:157–166
Swartz WJ, Schuetz AW (1975) Morphological diversity of oocytes released from the adult mouse ovary (1). Am J Anat 144:365–372
Wassarman PM, Josefowicz WJ, Letourneau GE (1976) Meiotic maturation of mouse oocytes in vitro: inhibition of maturation at specific stages of nuclear progression. J Cell Sci 22:331–342
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kim, H., Schuetz, A.W. Regulation of nuclear membrane assembly and maintenance during in vitro maturation of mouse oocytes: role of pyruvate and protein synthesis. Cell Tissue Res 265, 105–112 (1991). https://doi.org/10.1007/BF00318144
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00318144