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Activation of maturation promoting factor in Bufo arenarum oocytes: injection of mature cytoplasm and germinal vesicle contents

Published online by Cambridge University Press:  01 November 2006

G. Sánchez Toranzo
Affiliation:
Departamento de Biología del Desarrollo, San Miguel de Tucumán, Argentina
F. Bonilla
Affiliation:
Departamento de Biología del Desarrollo, San Miguel de Tucumán, Argentina
L. Zelarayán
Affiliation:
Departamento de Biología del Desarrollo, San Miguel de Tucumán, Argentina
J. Oterino
Affiliation:
Departamento de Biología del Desarrollo, San Miguel de Tucumán, Argentina
M.I. Bühler*
Affiliation:
Departamento de Biología del Desarrollo, San Miguel de Tucumán, Argentina
*
All correspondence to: Dra. Marta I. Bühler, Departamento de Biología del Desarrollo, Chacabuco 461, 4000 San Miguel de Tucumán, Argentina. Fax: +54 381 4248025. e-mail: mbuhler@fbqf.unt.edu.ar

Summary

Although progesterone is the established maturation inducer in amphibians, Bufo arenarum oocytes obtained during the reproductive period (spring–summer) resume meiosis with no need of an exogenous hormonal stimulus if deprived of their enveloping follicle cells, a phenomenon called spontaneous maturation. In this species it is possible to obtain oocytes competent and incompetent to undergo spontaneous maturation according to the seasonal period in which animals are captured. Reinitiation of meiosis is regulated by maturation promoting factor (MPF), a complex of the cyclin-dependent kinase p34cdc2 and cyclin B. Although the function and molecule of MPF are common among species, the formation and activation mechanisms of MPF differ according to species. This study was undertaken to evaluate the presence of pre-MPF in Bufo arenarum oocytes incompetent to mature spontaneously and the effect of the injection of mature cytoplasm or germinal vesicle contents on the resumption of meiosis. The results of our treatment of Bufo arenarum immature oocytes incompetent to mature spontaneously with sodium metavanadate (NaVO3) and dexamethasone (DEX) indicates that these oocytes have a pre-MPF, which activates and induces germinal vesicle breakdown (GVBD) by dephosphorylation on Thr-14/Tyr-15 by cdc25 phosphatase and without cyclin B synthesis. The injection of cytoplasm containing active MPF is sufficient to activate an amplification loop that requires the activation of cdc25 and protein kinase C, the decrease in cAMP levels, and is independent of protein synthesis. However, the injection of germinal vesicle content also induces GVBD in the immature receptor oocyte, a process dependent on protein synthesis but not on cdc25 phosphatase or PKC activity.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2006

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