The release from metaphase arrest in blue mussel oocytes.
In Mytilus edulis, shed oocytes are arrested at metaphase I of meiosis until fertilization. We previously suggested (Dubé and Dufresne, J. Exp. Zool. 256:323-332, 1990) that such a metaphase arrest depends upon a continuous synthesis of short-lived proteins, the destruction of which is sufficient to induce meiosis resumption. We further investigated the mechanism of metaphase release in blue mussel oocytes as triggered either by fertilization or by inhibition of protein synthesis (emetine) or phosphorylation (6-dimethylaminopurine, 6-DMAP). Treatment of unfertilized oocytes (UF) with emetine induces completion of the first meiotic cycle including extrusion of the polar body, followed by chromosome decondensation and by the formation of large membrane-bound nuclei, as visualized by Hoechst staining and transmission electron microscopy (TEM). Inhibition of protein phosphorylation with 6-DMAP induces directly chromosome decondensation and the formation of multiple nuclei surrounded by nuclear membrane. These interphasic nuclei exhibit continuous 3H-thymidine incorporation. p13 precipitation of p34 and associated proteins reveals "putative" cyclins in UF, no longer detected after metaphase/anaphase transition due to fertilization or emetine treatment. In the presence of 6-DMAP, new migrating forms are observed. The phosphorylated p34cdc2 homolog becomes dephosphorylated after fertilization or emetine treatment, whereas 6-DMAP induces its phosphorylation on tyrosine. Histone H1 kinase activity is reduced after these treatments, compared to the UF sample. Our results suggest that the metaphase/anaphase transition triggered by fertilization in blue mussel oocytes is induced by the rapid destruction of a set of continuously synthesized proteins accompanied by decreased histone H1 kinase activity. These events can be mimicked by inhibiting protein synthesis. Inhibition of protein phosphorylation would drive the cell to interphase without commitment to meiosis I.
Départment d'Océanographie, Université du Québec à Rimouski, Canada.
I
I
Néant
L
L
Dufresne
J
J
Morasse
C
C
Gicquaud
P
P
Guerrier
F
F
Dubé
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D049872:Bivalvia / Q000166:cytology; D016203:CDC2 Protein Kinase / Q000096:biosynthesis / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D004640:Emetine / Q000494:pharmacology; D005306:Fertilization; D008540:Meiosis; D008677:Metaphase / Q000187:drug effects / Q000502:physiology*; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D010766:Phosphorylation; D006656:Protamine Kinase / Q000378:metabolism; D014176:Protein Biosynthesis; D047428:Protein Kinase Inhibitors
D047428:Protein Kinase Inhibitors; C001020:N(6),N(6)-dimethyladenine; D004247:DNA; D006656:Protamine Kinase; D016203:CDC2 Protein Kinase; D000225:Adenine; D004640:Emetine