title: Morphology, sex ratio and gene expression of day 14 in vivo and in vitro bovine embryos.
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±0.33mm, 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.
authors.affiliation: Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology, Parque Estação Biológica, Av. W5 Norte final, Caixa Postal 02372, Brasília, DF 70770-917, Brazil.
authors.forename: Grazieli M
authors.initials: GM
authors.lastname: Machado
authors.forename: Allice R
authors.initials: AR
authors.lastname: Ferreira
authors.forename: Monique M
authors.initials: MM
authors.lastname: Guardieiro
authors.forename: Michele R
authors.initials: MR
authors.lastname: Bastos
authors.forename: José O
authors.initials: JO
authors.lastname: Carvalho
authors.forename: Carolina M
authors.initials: CM
authors.lastname: Lucci
authors.forename: Tiago O
authors.initials: TO
authors.lastname: Diesel
authors.forename: Roberto
authors.initials: R
authors.lastname: Sartori
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Rumpf
authors.forename: Maurício M
authors.initials: MM
authors.lastname: Franco
authors.forename: Margot A N
authors.initials: MA
authors.lastname: Dode
mesh_terms: D000367:Age Factors; D000818:Animals; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D046149:Embryo Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology* / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D051272:Glucose Transporter Type 1 / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012732:Sex Determination Analysis; D012744:Sex Ratio; D018709:Statistics, Nonparametric; D016019:Survival Analysis
chemical_list: D017931:DNA Primers; D051272:Glucose Transporter Type 1; D018840:HSP70 Heat-Shock Proteins