Activity of maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK) after parthenogenetic activation of ovine oocytes.
The maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK) are the key regulators of both meiotic and mitotic cell cycles. Knowledge of the dynamics of these two kinases during the transition from meiosis to mitosis would be of great importance for cloning by nuclear transfer. In this study, experiments were designed to assay the changes of MPF and MAP kinase activity of in vitro matured ovine oocytes after chemical activation and culture in 0 mM or 2 mM 6-dimethylaminopurine (6-DMAP) for 12 h. Moreover, to determine the biological significance of the fluctuations of MPF, activated oocytes were fused with GV-staged partners. The biochemical results showed that the high MPF activity of MII oocytes fell to basal level precipitously within the first hour after activation, started to increase at 6-8 h, rising to 80 +/- 4% of MII after 12 h. MAPK activity decreased to a low level 4 h after activation, increased between 6-12 h, but remained below 30 +/- 3.6% of MII values. The incubation with 6-DMAP had no effect on the kinetics of MPF and MAP kinase activity. Fusion of MII oocytes to GV partners induced rapid breakdown of the GV, whereas no breakdown occurred when GV were fused with eggs in the first hours post activation. Interestingly, the high biochemical levels of MPF activity at 8-12 h after activation were not able to induce GVBD in fusion partners.
Bogliolo
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Obstetrics Section of the Institute of General Pathology, Pathological Anatomy and Veterinary Obstetrics-Surgery Clinic, University of Sassari, Sassari, Italy. vetfis@ssmain.uniss.it
Ledda
S
S
Leoni
G
G
Naitana
S
S
Moor
R M
RM
D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle; D002459:Cell Fusion; D019976:Cloning, Organism / Q000379:methods*; D004789:Enzyme Activation; D015151:Immunoblotting; D007700:Kinetics; D020935:MAP Kinase Signaling System; D016200:Maturation-Promoting Factor / Q000378:metabolism*; D008954:Models, Biological; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D010312:Parthenogenesis; D010766:Phosphorylation; D012756:Sheep; D013997:Time Factors
C001020:N(6),N(6)-dimethyladenine; D016200:Maturation-Promoting Factor; D000225:Adenine