Abstract
High hydrostatic pressure (HHP) has been used to pre-condition embryos before essential, yet potentially detrimental procedures such as cryopreservation. However, the mechanisms for HHP are poorly understood. We treated bovine blastocysts with three different HHP (40, 60 and 80 MPa) in combination with three recovery periods (0, 1 h, 2 h post HHP). Re-expansion rates were significantly higher at 40 and 60 but lower at 80 MPa after vitrification-warming in the treated groups than controls. Microarray analysis revealed 399 differentially expressed transcripts, representing 254 unique genes, among different groups. Gene ontology analysis indicated that HHP at 40 and 60 MPa promoted embryo competence through down-regulation of genes in cell death and apoptosis, and up-regulation of genes in RNA processing, cellular growth and proliferation. In contrast, 80 MPa up-regulated genes in apoptosis, and down-regulated protein folding and cell cycle-related genes. Moreover, gene expression was also influenced by the length of the recovery time after HHP. The significantly over-represented categories were apoptosis and cell death in the 1 h group, and protein folding, response to unfolded protein and cell cycle in the 2 h group compared to 0 h. Taken together, HHP promotes competence of vitrified bovine blastocysts through modest transcriptional changes.
Highlights
Gamma growth arrest and DNA-damage-inducible (GADD45G) were found to be significantly up-regulated by High hydrostatic pressure (HHP) treatment[12]
The aim of the present study was to evaluate the effects of HHP treatments at three different levels with two different recovery times on the gene expression of bovine in vitro produced (IVP) vitrified blastocysts
Re-expansion rates were significantly affected by HHP and recovery time, but not by temperature
Summary
Gamma growth arrest and DNA-damage-inducible (GADD45G) were found to be significantly up-regulated by HHP treatment[12]. In bovine blastocysts subjected to HHP treatment, several candidate stress genes have been examined These include stress related genes SOD2, glutathione peroxidase 4 (GPX4) and heat shock 70 kDa protein 1 A (HSPA1A). Cumulative analysis of these genes revealed a similar pattern of expression, with a tendency for peak transcript abundance 1 h after HHP treatment[14]. The aim of the present study was to evaluate the effects of HHP treatments at three different levels with two different recovery times on the gene expression of bovine in vitro produced (IVP) vitrified blastocysts To our knowledge, this is the first report of transcriptional profiling of bovine blastocysts treated by HHP. Protein folding, cell cycle regulation, RNA processing and translation were found to be affected by HHP
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