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Partial inhibition of nitric oxide synthase activity stimulates the nuclear maturation progression of bovine cumulus–oocyte complex in vitro in the presence of hemisections of the follicular walls

Published online by Cambridge University Press:  15 May 2020

Diego Fernando Dubeibe
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
Maria Clara Caldas-Bussiere*
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
Valter Luiz Maciel Jr
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
Wlaisa Sampaio
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
Paulo B.D. Gonçalves
Affiliation:
Laboratório de Biotecnologia e Reprodução Animal, Universidade Federal de Santa Maria – UFSM, Santa Maria, Rio Grande do Sul-RS, Cep 97105-900, Brazil.
Matheus P. De Cesaro
Affiliation:
Laboratório de Biotecnologia e Reprodução Animal, Universidade Federal de Santa Maria – UFSM, Santa Maria, Rio Grande do Sul-RS, Cep 97105-900, Brazil.
Celia Raquel Quirino
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
Márcia R. Faes
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
Carla S. Paes de Carvalho
Affiliation:
Laboratório de Reprodução e Melhoramento Genético Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro – RJ, Cep 28013-602, Brazil.
*
Author for correspondence: Maria Clara Caldas-Bussiere. Laboratório de Reprodução e Melhoramento Genético Animal, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, Rio de Janeiro-RJ, Cep 28013-602, Brazil. Tel: +55 22 27397285. E-mail: [email protected]

Summary

This study aimed to assess the effects of the inhibition of nitric oxide synthase (NOS) on events that modulate bovine in vitro oocyte maturation. Cumulus–oocyte complexes (COCs) were cultured with hemisections (HSs) of the follicular walls in a maturation medium supplemented with different concentrations (0.1–10.0 mM) of Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME). Controls consisted of COCs cultured in the presence (+HSs) or absence of HSs (–HSs) with no additional l-NAME supplementation. The following parameters were assessed: oocyte nuclear maturation stage; cumulus cell (CC) membrane integrity; nitrate/nitrite, progesterone, and estradiol concentrations in the culture medium at 22 h of cultivation; and the concentrations of cGMP and cAMP in COCs during the first hour of maturation. The addition of 1.0 mM l-NAME increased the percentage of oocytes that reached metaphase II (MII) and the percentage of intact CCs (P < 0.05). All l-NAME concentrations reduced the nitrate/nitrite concentrations (P < 0.05), but none affected steroid concentrations compared with control +HSs (P > 0.05). The addition of 1.0 mM l-NAME reduced cGMP concentrations at 3 h and increased cAMP concentrations in the first hour of culture (P < 0.05). Our findings suggest that the NOS/NO/cGMP pathway participates in meiosis progression (MI to MII) of the bovine oocytes matured in vitro in the presence of hemisections of the follicular walls. Lastly, the mechanisms that lead to the progression of meiosis after NOS inhibition do not involve changes in steroid production.

Type
Research Article
Copyright
© The Author(s), 2020. Published by Cambridge University Press

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