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8 February 2021 Activation of bovine oocytes by protein synthesis inhibitors: new findings on the role of MPF/MAPKs
Cecilia Valencia, Felipe Alonso Pérez, Carola Matus, Ricardo Felmer, María Elena Arias
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Abstract

The present study evaluated the mechanism by which protein synthesis inhibitors activate bovine oocytes. The aim was to analyze the dynamics of MPF and MAPKs. MII oocytes were activated with ionomycin (Io), ionomycin+anisomycin (ANY) and ionomycin+cycloheximide (CHX) and by in vitro fertilization (IVF). The expression of cyclin B1, p-CDK1, p-ERK1/2, p-JNK, and p-P38 were evaluated by immunodetection and the kinase activity of ERK1/2 was measured by enzyme assay. Evaluations at 1, 4, and 15 hours postactivation (hpa) showed that the expression of cyclin B1 was not modified by the treatments. ANY inactivated MPF by p-CDK1Thr14-Tyr15 at 4 hpa (P < 0.05), CHX increased pre-MPF (p-CDK1Thr161 and p-CDK1Thr14-Tyr15) at 1 hpa and IVF increased p-CDK1Thr14-Tyr15 at 17 hours postfertilization (hpf) (P < 0.05). ANY and CHX reduced the levels of p-ERK1/2 at 4 hpa (P < 0.05) and its activity at 4 and 1 hpa, respectively (P < 0.05). Meanwhile, IVF increased p-ERK1/2 at 6 hpf (P < 0.05); however, its kinase activity decreased at 6 hpf (P < 0.05). p-JNK in ANY, CHX, and IVF oocytes decreased at 4 hpa (P < 0.05). p-P38 was only observed at 1 hpa, with no differences between treatments. In conclusion, activation of bovine oocytes by ANY, CHX, and IVF inactivates MPF by CDK1-dependent specific phosphorylation without cyclin B1 degradation. ANY or CHX promoted this inactivation, which seemed to be more delayed in the physiological activation (IVF). Both inhibitors modulated MPF activity via an ERK1/2-independent pathway, whereas IVF activated the bovine oocytes via an ERK1/2-dependent pathway. Finally, ANY does not activate the JNK and P38 kinase pathways.

Summary sentence

Activation of bovine oocytes by protein synthesis inhibitors modulate MPF by CDK1-dependent specific phosphorylation and via an MAPKs-independent pathway.

© The Author(s) 2021. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
Cecilia Valencia, Felipe Alonso Pérez, Carola Matus, Ricardo Felmer, and María Elena Arias "Activation of bovine oocytes by protein synthesis inhibitors: new findings on the role of MPF/MAPKs," Biology of Reproduction 104(5), 1126-1138, (8 February 2021). https://doi.org/10.1093/biolre/ioab019
Received: 16 December 2020; Accepted: 2 February 2021; Published: 8 February 2021
KEYWORDS
anisomycin
CDK1
cyclinB1
cycloheximide
Erk1/2
MAPKs
Oocytes activation
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