Project description:Changes in gene expression induced by the Cryotop vitrification procedure in bovine blastocysts using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos at the blastocyst stage (144 to 156 hours post insemination) were vitrified using the Cryotop system and compared with non-vitrified (control) embryos. After vitrification, the embryos were warmed and cultured for an additional 4 hours. Embryos that re-expanded or developed to the expanded blastocyst stage were used for microarray analysis.
Project description:Changes in gene expression induced by the Cryotop vitrification procedure in bovine blastocysts using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos at the blastocyst stage (144 to 156 hours post insemination) were vitrified using the Cryotop system and compared with non-vitrified (control) embryos. After vitrification, the embryos were warmed and cultured for an additional 4 hours. Embryos that re-expanded or developed to the expanded blastocyst stage were used for microarray analysis. Four pools of vitrified embryos were hybridized against four pools of control embryos in a dye-swap design.
Project description:Affymetrix Human GeneChips are used to profile gene expression of bovine tissues and embryos to identify uniquely expressed genes in bovine in-vitro fertilized embryos by comparing with seven bovine adult tissues through gene clustering
Project description:Embryo vitrification involves exposure to high concentrations of cryoprotectants and osmotic stress during cooling and warming. A multitude of these factors have the potential of affecting gene expression. In this study, in vitro produced bovine embryos at the blastocyst stage were submitted to vitrification. Four recipients each were used for transferring non-vitrified (80) and vitrified (80) embryos. Twelve non-vitrified and nine vitrified viable day-14 (D14) embryos were recovered by uterine flushing. RNA-seq analysis of whole embryo or isolated trophectoderm (TE) from vitrified and fresh recovered D14 embryos revealed a total of 927 genes changed their expression as a result of vitrification, among them 782 and 145 genes were up- and down-regulated, respectively. In TE isolates, vitrification resulted in 4,096 and 280 up- and down-regulated genes, respectively. In addition, we found 671 and 61 genes commonly up- and down-regulated in both vitrified whole embryo and TE. Commonly up-regulated pathways by vitrification included epithelial adherens junctions, sirtuin signaling, germ cell-sertoli cell junction, ATM signaling, NER, and protein ubiquitination pathways. The commonly down-regulated pathways included EIF2 signaling, oxidative phosphorylation, mitochondrial dysfunction, regulation of eIF4 and p70S6K signaling, mTOR signaling, sirtuin singling, and NER pathways. Our analysis identified specific pathways and implicated specific gene expression patterns important to cryopreservation affecting embryo developmental competence.
Project description:miRNA profile of the bovine pretransfer endometrium based on pregnancy success after in vivo and in vitro produced embryos transfer
Project description:In vitro maturation (IVM) of the oocytes is a routine method in bovine embryo production. The competence of bovine oocytes to develop into embryo after IVM and in vitro fertilization (IVF) is lower as compared to in vivo preovulatory oocytes. Cumulus cells (CC) that enclose an oocyte are involved in the acquisition of oocyte quality during maturation. Using transcriptomic approach we compared cumulus cells gene expression during IVM with that in vivo preovulatory period.
