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In vitro embryos production after oocytes treatment with forskolin

Published online by Cambridge University Press:  24 February 2015

Daniela Martins Paschoal ,
Rosiára Rosária Dias Maziero ,
Mateus José Sudano ,
Midyan Daroz Guastali ,
Luis Eduardo Vergara ,
Letícia Ferrari Crocomo ,
João Ferreira de Lima-Neto ,
Luis Carlos Oña Magalhães ,
Bianca Andriolo Monteiro  and
Tatiana da Silva Rascado
...Show all authors
Daniela Martins Paschoal*
Affiliation:
São Paulo State University (UNESP), School of Veterinary Medicine and Animal Science (FMVZ), Department of Animal Reproduction and Veterinary Radiology, Rubião Jr. s/n, Botucatu/SP, Brazil
Rosiára Rosária Dias Maziero
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Mateus José Sudano
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Midyan Daroz Guastali
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Luis Eduardo Vergara
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Letícia Ferrari Crocomo
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
João Ferreira de Lima-Neto
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Luis Carlos Oña Magalhães
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Bianca Andriolo Monteiro
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Tatiana da Silva Rascado
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
Alício Martins Jr
Affiliation:
Department of Clinical Surgery and Animal Reproduction, School of Veterinary Medicine (FMVA), São Paulo State University (UNESP), Araçatuba, SP, Brazil.
Claudia Lima Verde Leal
Affiliation:
Department of Basic Science, School of Animal Science and Food Engineering (FZEA), São Paulo University (USP), Pirassununga/SP, Brazil.
Fernanda da Cruz Landim-Alvarenga
Affiliation:
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science (FMVZ), São Paulo State University (UNESP), Botucatu, SP, Brazil.
*
All correspondence to: Daniela Martins Paschoal. São Paulo State University (UNESP), School of Veterinary Medicine and Animal Science (FMVZ), Department of Animal Reproduction and Veterinary Radiology, Rubião Jr. s/n, Botucatu/SP, Brazil, 18618–970. Tel:/Fax: +55 14 3880 2119. E-mail: [email protected]
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Summary

The inhibition of nuclear maturation allows time for the oocyte to accumulate molecules that are important for embryonic development. Thus, the objective of this work was to evaluate the effect of blocking oocyte meiosis with the addition of forskolin, an efficient inhibitor of nuclear maturation, in in vitro maturation (IVM) medium. Forskolin was added to the IVM medium for 6 h at concentrations of 0.1 mM, 0.05 mM or 0.025 mM, then the oocytes were allowed to mature in drug-free medium for 18 h. The oocytes were assessed for the stage of nuclear maturation, the activity and distribution of mitochondria, oocyte ultrastructure, the number of viable cells and the apoptosis rate. After forskolin treatment, the oocytes were fertilized in vitro and cultured for 7 days. On day 7, the blastocyst rate, the ultrastructure, the number of intact cells and the apoptosis rate of the blastocysts were measured. No differences were observed for the stage of nuclear maturation of the oocyte, the mitochondrial activity and distribution, the blastocyst rate or total number of intact cells. However, a higher rate of apoptosis was observed in the blastocysts produced from oocytes blocked for 6 h with the higher concentration of forskolin (P < 0.05). We conclude that all the experimental groups reached the MII stage after the addition of forskolin and that the highest concentration of forskolin caused cellular degeneration without harming embryo production on the 7th day.

Keywords

ApoptosiscAMPForskolinIVMMeiosis
Type
Research Article
Information
Zygote , Volume 24 , Issue 2 , April 2016 , pp. 161 - 171
Copyright
Copyright © Cambridge University Press 2015 

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