title: Effect of vitrification using the Cryotop method on the gene expression profile of in vitro-produced bovine embryos.
abstract: The present study analyzed the changes in gene expression induced by the Cryotop vitrification technique in bovine blastocyst-stage embryos, using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos were vitrified and compared with nonvitrified (control) embryos. After vitrification, embryos were warmed and cultured for an additional 4 hours. Survived embryos were used for microarray analysis and quantitative polymerase chain reaction (qPCR) quantification. Survival rates were higher (P < 0.05) in the control embryos (100%) than in the vitrified embryos (87%). Global gene expression analysis revealed that only 43 out of 21,139 genes exhibited significantly altered expression in the vitrified embryos compared to the control embryos, with a very limited fold change (P < 0.05). A total of 10 genes were assessed by qPCR. Only the FOS-like antigen 1 (FOSL1) gene presented differential expression (P < 0.05) according to both the array and qPCR methods, and it was overexpressed in vitrified embryos. Although, the major consequence of vitrification seems to be the activation of the apoptosis pathway in some cells. Indeed, FOSL1 is part of the activating protein 1 transcription factor complex and is implicated in a variety of cellular processes, including proliferation, differentiation, and apoptosis. Therefore, our results suggest that a limited increase in the rate of apoptosis was the only detectable response of the embryos to vitrification stress.
authors.affiliation: School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, Federal District, Brazil.
authors.forename: Ligiane
authors.initials: L
authors.lastname: de Oliveira Leme
authors.affiliation: Centre de Recherche en Biologie de la Reproduction, Faculté des Sciences de l'Agriculture et de l'Alimentation, Département des Sciences Animales, Pavillon INAF, Université Laval, Québec City, Quebec, Canada.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Dufort
authors.affiliation: School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, Federal District, Brazil.
authors.forename: José Felipe Warmling
authors.initials: JF
authors.lastname: Spricigo
authors.affiliation: School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, Federal District, Brazil.
authors.forename: Thiago Felipe
authors.initials: TF
authors.lastname: Braga
authors.affiliation: Centre de Recherche en Biologie de la Reproduction, Faculté des Sciences de l'Agriculture et de l'Alimentation, Département des Sciences Animales, Pavillon INAF, Université Laval, Québec City, Quebec, Canada.
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Sirard
authors.affiliation: Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Brasília, Federal District, Brazil.
authors.forename: Maurício Machaim
authors.initials: MM
authors.lastname: Franco
authors.affiliation: School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, Federal District, Brazil; Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Brasília, Federal District, Brazil. Electronic address: margot.dode@embrapa.br.
authors.forename: Margot Alves Nunes
authors.initials: MA
authors.lastname: Dode
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000196:embryology*; D015925:Cryopreservation / Q000662:veterinary*; D046149:Embryo Culture Techniques; D005307:Fertilization in Vitro / Q000662:veterinary*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D040081:Protein Array Analysis / Q000662:veterinary; D059467:Transcriptome; D058989:Vitrification
keywords: Cattle; Cryotop; Embryo; Gene expression; Microarray; Transcriptome; Vitrification