Jiang Zongliang, Harrington Patrick, Zhang Ming, Marjani Sadie L, Park Joonghoon, Kuo Lynn, Pribenszky Csaba, Tian Xiuchun Cindy
Center for Regenerative Biology, Department of Animal Science, University of Connecticcut, Storrs, Connecticut, 06269, USA.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Animal Reproduction Institute, Guangxi University, Nanning, Guangxi, China, 510005.
Sci Rep. 2016 Feb 17;6:21215. doi: 10.1038/srep21215.
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.
高静水压（HHP）已被用于在进行诸如冷冻保存等必要但可能有害的操作之前对胚胎进行预处理。然而，人们对HHP的机制了解甚少。我们用三种不同的高静水压（40、60和80兆帕）处理牛囊胚，并结合三个恢复期（高静水压处理后0、1、2小时）。处理组玻璃化冷冻解冻后的再扩张率在40和60兆帕时显著高于对照组，但在80兆帕时低于对照组。微阵列分析显示不同组之间有399个差异表达的转录本，代表254个独特基因。基因本体分析表明，40和60兆帕的高静水压通过下调细胞死亡和凋亡相关基因以及上调RNA加工、细胞生长和增殖相关基因来促进胚胎发育能力。相比之下，80兆帕上调了凋亡相关基因，并下调了蛋白质折叠和细胞周期相关基因。此外，基因表达也受高静水压处理后恢复时间长短的影响。与0小时相比，1小时组中显著富集的类别是凋亡和细胞死亡，2小时组中是蛋白质折叠、对未折叠蛋白的反应和细胞周期。综上所述，高静水压通过适度的转录变化促进玻璃化牛囊胚的发育能力。
