State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, China.
Reprod Biomed Online. 2013 May;26(5):491-9. doi: 10.1016/j.rbmo.2013.01.005. Epub 2013 Jan 19.
Previous studies by this study group have showed that heat shock protein 27 (Hsp27), expressed in the oocyte of growing follicles, is down-regulated in polycystic ovary syndrome ovaries and that down-regulation of Hsp27 improves the maturation of mouse oocytes and increases early apoptosis of oocytes. In this study, the effect of Hsp27 on early embryo development in the mouse was observed. Following microinjection of AdCMV-Hsp27 or AdsiRNA-Hsp27 into the cytoplasm of mouse zygotes, blastocyst morphology was observed and cell apoptosis of blastocysts was detected by TUNEL. After culture in vitro for 96h, blastocysts were analysed for Hsp27 expression by real-time PCR and immunofluorescence. The blastocyst formation rate and embryo quality were evaluated. The expression of Hsp27 was significantly increased in embryos with Hsp27 overexpression (AdCMV-Hsp27), while it was significantly suppressed by 75% in embryos with the gene silenced (AdsiRNA-Hsp27; both P<0.05). Cell apoptosis in blastocysts, blastocyst formation rate and embryo quality were unaffected by Hsp27 overexpression or gene silencing. In conclusion, overexpression or down-regulation of Hsp27 in zygotes, as a single factor, does not significantly affect the subsequent embryonic development.
本研究小组的先前研究表明,在生长卵泡的卵母细胞中表达的热休克蛋白 27(Hsp27)在多囊卵巢综合征卵巢中下调,并且 Hsp27 的下调可改善小鼠卵母细胞的成熟并增加卵母细胞的早期凋亡。在这项研究中,观察了 Hsp27 对小鼠早期胚胎发育的影响。将 AdCMV-Hsp27 或 AdsiRNA-Hsp27 微注射到小鼠受精卵的细胞质中后,观察囊胚形态,并通过 TUNEL 检测囊胚的细胞凋亡。体外培养 96h 后,通过实时 PCR 和免疫荧光法分析囊胚中的 Hsp27 表达。评估囊胚形成率和胚胎质量。Hsp27 过表达(AdCMV-Hsp27)的胚胎中 Hsp27 的表达明显增加,而基因沉默(AdsiRNA-Hsp27)的胚胎中 Hsp27 的表达则明显抑制(均 P<0.05)。囊胚中的细胞凋亡,囊胚形成率和胚胎质量不受 Hsp27 过表达或基因沉默的影响。总之,作为单一因素,受精卵中 Hsp27 的过表达或下调不会显著影响随后的胚胎发育。