1 东莞市人民医院生殖医学科，广东 东莞 523059
2 东莞职业技术学院卫生健康学院，广东 东莞 523186
目的：探究小鼠卵母细胞miR-135的表达对胚胎发育的影响。方法：实时荧光定量PCR(real-time polymerase chain reaction，real-time PCR)检测小鼠不同发育质量囊胚中miR-135的表达。小鼠卵母细胞注射miR-135抑制剂(miR-135 inhibitor)后，再体外受精(in vitro fertilization，IVF)，分析不同质量囊胚构成比；检测植入前胚胎发育过程中2细胞期、4细胞期、桑椹胚和囊胚的数量；使用荧光染色观察囊胚中囊胚中胰岛素样生长因子1受体(insulin-like growth factor 1 receptor，IGF1R)的表达及分布；将囊胚植入小鼠后，观察发育后胎儿的冠臀长度。结果：miR-135在高质量发育的囊胚中高表达。卵母细胞注射miR-135抑制剂后，再进行IVF，高质量囊胚比例降低和植入前胚胎发育迟缓；显微镜下观察发现：囊胚中IGF1R的表达无明显改变，但分布模式出现变化，表达集中于基底膜上；植入后胎鼠的冠臀长度明显变短。结论：抑制小鼠卵母细胞miR-135的表达会抑制胚胎发育。
Effect of miR-135 expression on embryonic development in mouse oocytes
CorrespondingAuthor: LIANG Jiandi
This work was supported by Dongguan Medical and Health Technology Program, Guangdong Province
Objective: To investigate the effects of miR-135 expression in oocytes on embryonic development. Methods: Real-time quantitative polymerase chain reaction (real-time PCR) was used to detect the expression of miR-135 in blastocysts with different developmental quality. Then, the oocytes were injected with miR-135 inhibitor before in vitro fertilization, the composition ratio of different developmental quality was analyzed. The number of 2 cell stage, 4 cell stage, morula stage and blastocyst stage during the development of preimplantation embryos was detected. The expression and distribution of IGF1R in blastocysts was observed by fluorescence staining. After the blastocyst was implanted into mice, the length of the crown and hip of the developing fetus was observed. Results: MiR-135 was highly expressed in high quality blastocysts. After the oocytes were injected with miR-135 inhibitor and then fertilized in vitro, the proportion of high-quality blastocysts was decreased and preimplantation embryonic development became retardation. Microscopic observation showed no significant change in the expression of IGF1R in blastocysts, but the distribution pattern was changed, and the expression was concentrated on the basement membrane. After implantation, the length of the crown and buttock in fetal mice became shorter. Conclusion: Inhibition of miR-135 expression in mouse oocytes inhibits embryonic development.
miR-135; oocyte; embryo development; developmental biology