PLCζ对小鼠圆形精子细胞注射后卵母细胞受精及胚胎发育的影响

Effects of PLCζ injection into mouse round spermatid on the oocyte fertilization and embryo development

目的探讨PLCζ作为激活剂的可行性及对ROSI后卵母细胞受精及胚胎发育的影响。方法在体外构建重组质粒pCRII-TOPO-PLCζ,诱导表达后采用质谱鉴定和Western blot检测抗原性。重组PLCζ蛋白作为小鼠ROSI卵母细胞的激活剂,记录受精及胚胎发育情况并统计分析。结果成功构建了重组质粒pCRII-TOPO-PLCζ,体外表达后经质谱鉴定为小鼠PLCζ蛋白,并具有良好的抗原性。重组PLCζ蛋白激活卵母细胞后的受精率、2-4细胞率、8细胞率及成囊率与空白组之间的差异均无显著性(P>0.05)。结论本研究中重组PLCζ蛋白对卵母细胞受精及胚胎发育无显著影响,作为小鼠ROSI后的卵母细胞激活剂的可行性值得商榷。

Objective To investigate the feasibility of PLCζas an activator and its effect on fertilization and embryo development for oocytes after mouse round spermatid injection.Methods The recombinant plasmid pCRII-TOPOPLCζwas constructed in vitro and its expression was induced.The protein was identified by mass spectrometry and the antigenicity was detected by Western blot.Recombinant PLCζprotein was used as an activator of mouse ROSI oocytes,and fertilization and embryo development were recorded and analyzed statistically.Results The recombinant plasmid pCRIITOPO-PLCζwas successfully constructed,and the recombinant PLCζprotein was identified by mass spectrometry and had good antigenicity.The fertilization rate,two-to four-cell rate,eight-cell rate,and cystic rate associated with the recombinant PLCζprotein did not differ significantly from those in the blank group(P>0.05).Conclusions The recombinant PLCζprotein has no significant effect on oocyte fertilization and embryo development in this study.The feasibility of being an oocyte activator after mouse ROSI is questionable.