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Meiotic arrest as an alternative to increase the production of bovine embryos by somatic cell nuclear transfer

Published online by Cambridge University Press:  26 October 2016

F.M.C. Caixeta
Affiliation:
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília-DF, Brazil.
R.V. Sousa
Affiliation:
EMBRAPA Genetic Resources and Biotechnology, Brasília-DF, Brazil.
A.L. Guimarães
Affiliation:
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília-DF, Brazil.
L.O. Leme
Affiliation:
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília-DF, Brazil.
J.F.W. Sprícigo
Affiliation:
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília-DF, Brazil.
S.B. Senna Netto
Affiliation:
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília-DF, Brazil.
I. Pivato
Affiliation:
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília-DF, Brazil.
M.A.N. Dode*
Affiliation:
EMBRAPA Genetic Resources and Biotechnology, Parque Estação Biológica, Final Av. W5/N, Prédio PBI, 70770-900, Brasília-DF, Brazil. EMBRAPA Genetic Resources and Biotechnology, Brasília-DF, Brazil.
*
All correspondence to: Margot Alves Nunes Dode, EMBRAPA Genetic Resources and Biotechnology, Parque Estação Biológica, Final Av. W5/N, Prédio PBI, 70770–900, Brasília-DF, Brazil. Tel: +55 61 34484659. Fax: +55 61 3340 3658. E-mail: margot.dode@embrapa.br

Summary

This study aimed to evaluate the effect of meiotic arrest using phosphodiesterase type 3A (PDE 3A) inhibitors, cilostamide and C-type natriuretic peptide (NPPC), on pre-maturation (PM) of oocytes to be used in the production of cloned embryos. Nuclear maturation, in vitro embryo production (IVP), somatic cell nuclear transfer (SCNT) and parthenogenetic activation (PA), and total cells number of cloned embryos were evaluated. The results were analysed by chi-squared and Kruskal–Wallis test with a P-value <0.05 considered to be significant. Approximately 87.8% of the oocytes remained at germinal vesicle stage (GV) after 6 h of PM with 5 μM of cilostamide, confirming the meiotic block. Embryo development in IVP was similar (P > 0.05) between control and PM, both for cleavage (78.2% and 76.9%) and blastocyst (35.5% and 29.3%) rates. After SCNT, cleavage rate was also similar (P > 0.05) between control and PM (66% and 51.9%) however, blastocyst rate was lower (P < 0.05) in the PM group than in the control group (7.4% and 30.2%). After 6 h of PM with 100 nM of NPPC, approximately 84.9% of the oocytes remained at GV. No difference was found between control and PM in cleavage (69.2% and 76.1%) and blastocyst rates (37,4% and 35%) after IVP. Similarly, no differences between PM and control groups were observed for cleavage (69.2% and 68.4%) and blastocyst (24.4% and 21.5%) rates. SCNT and PA embryos from control or PM oocytes had similar total cell number. It can be concluded that PM for 6 h with 100 nM NPPC is feasible for cloned embryo production without affecting embryo outcome.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2016 

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