Morphology, sex ratio and gene expression of Day 14 in vivo and in vitro bovine embryos
Grazieli M. Machado A B , Allice R. Ferreira A C , Monique M. Guardieiro D , Michele R. Bastos C , José O. Carvalho A D , Carolina M. Lucci E , Tiago O. Diesel A B , Roberto Sartori D , Rodolfo Rumpf A , Maurício M. Franco A and Margot A. N. Dode A B F
+ Author Affiliations
- Author Affiliations
A Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology, Parque Estação Biológica, Av. W5 Norte (final), Caixa Postal 02372, Brasília, DF 70770-917, Brazil.
B College of Agriculture and Veterinary, University of Brasília, Campus Universitário Darcy Ribeiro, Asa Norte, Instituto Central de Ciências Ala Sul, Caixa Postal 4508, Brasília, DF 70910-970, Brazil.
C College of Veterinary and Animal Science, São Paulo State University, Distrito Rubião Júnior, Botucatu, SP 18618-970, Brazil.
D Department of Animal Science, College of Agriculture Luiz de Queiroz (ESALQ), University of São Paulo, Piracicaba, SP 13418-900, Brazil.
E Department of Physiological Science, Institue of Biological Science, University of Brasília, Campus Universitário Darcy Ribeiro, Asa Norte, Brasília, DF 70910-970, Brazil.
F Corresponding author. Email: margot@cenargen.embrapa.br
Reproduction, Fertility and Development 25(4) 600-608 https://doi.org/10.1071/RD11282
Submitted: 5 November 2011 Accepted: 5 May 2012 Published: 7 June 2012
Abstract
The present study was designed to compare Day 14 bovine embryos that were produced entirely in vitro using the post-hatching development (PHD) system with in vivo-derived embryos without or with transient PHD culture from Day 7 to Day 14. Embryos on Day 14 were used for sex determination and gene expression analysis of PLAC8, KRT8, CD9, SLC2A1, SLC2A3, PGK1, HSF1, MNSOD, HSP70 and IFNT using real-time quantitative (q) polymerase chain reaction (PCR). First, Day 7 in vivo- and in vitro-produced embryos were subjected to the PHD system. A higher rate of survival was observed for in vitro embryos on Day 14. Comparing Day 14 embryos produced completely in vivo or completely in vitro revealed that the mean size of the former group was greater than that of the latter (10.29 ± 1.83 vs 2.68 ± 0.33 mm, respectively). Expression of the HSP70 and SLC2A1 genes was down- and upregulated, respectively, in the in vitro embryos. The present study shows that in vitro embryos cultured in the PHD system are smaller than in vivo embryos and that of the 10 genes analysed, only two were differentially expressed between the two groups. These findings indicate that, owing to the poor survival rate, the PHD system is not reliable for evaluation of in vitro embryo quality.
Additional keywords: agarose tunnels, post-hatching embryos.
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