Project description: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, 1h, 2h 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 1h group, and protein folding, response to unfolded protein and cell cycle in the 2h group compared to 0h. Taken together, HHP promotes competence of vitrified bovine blastocysts through modest transcriptional changes.
Project description: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, 1h, 2h 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 1h group, and protein folding, response to unfolded protein and cell cycle in the 2h group compared to 0h. Taken together, HHP promotes competence of vitrified bovine blastocysts through modest transcriptional changes. 4*3*2 design experiment. The following treatments were included: (1) Control embryos were left untreated in the incubator (one atmospheric pressure or 0.1 MPa); (2) treatment groups were assigned to 40, 60 and 80 MPa HHP for 1h at either 24°C (room temperature) or 39°C (body temperature), followed by three different recovery time periods (0, 1 and 2h) post-HHP in the holding medium. Biological replicates: 3 control replicates. Technical replicates: dye-swap.
Project description:Vegetative cells of B. subtilis can recover from injury caused by high hydrostatic pressure (HHP) treatment at 250 MPa. DNA microarray analysis revealed that many ribosomal genes and translation relating factors (e.g. translation initiation factors) were induced during a growth-arrested phase after the HHP treatment. Expression of cold shock-responsive genes, whose products play key roles for efficient translation, and heat shock-responsive genes, whose product mediates the correct protein folding or degrades misfolded proteins, were also induced. In contrast, the expression of hpf, whose product (Hpf) is involved in ribosome inactivation through dimerization of 70S ribosomes, was repressed. Sucrose density gradient sedimentation analysis showed that ribosomes were dissociated in a pressure-dependent manner and then reconstructed during the growth-arrested phase. These results suggested that the translational machinery can be preferentially reconstructed in the HHP-injured cells. We also found that cell growth after HHP-induced injury was apparently inhibited by Mn2+ or Zn2+ supplemented to the recovery medium. Ribosome reconstruction in HHP-injured cells was significantly delayed in the presence of Mn2+ or Zn2+. Moreover, Zn2+ but not Mn2+ stimulated dimer formation of ribosomes in HHP-injured cells. This Zn2+-dependent accumulation of ribosome dimer was no longer observed in a Δhpf mutant lacking the functional Hpf. Furthermore, the growth recovery of the Δhpf mutant in the Zn2+-supplemented medium was faster than that of the parent strain. Thus, our results indicate that Zn2+ can prevent ribosome reconstruction by stimulating the Hpf-dependent ribosome dimerization, thereby inhibiting the growth recovery of the HHP-injured B. subtilis cells.
2019-10-26 | GSE126203 | GEO
Project description:Microbial community of soil samples during anoxic incubation under different kinds of sterilization
| PRJNA949398 | ENA
Project description:Shotgun metagenomics of jeevamrit formulation under different incubation
Project description:Transcription profiling of mouse oocytes treated with 20 MPa hydrostatic pressure for 60 minutes at 37 °C comparing control oocytes kept under identical conditions as pressure treated ones, except HHP treatment.
2012-01-06 | GSE28410 | GEO
Project description:Microbial analysis under different remediation measures and incubation experiments