未受精人卵母细胞及废弃胚胎线粒体基因的表达  

Mitochondrial Gene Expression in Human Unfertilized Oocytes and Cleavage-stage Embryos

作　　者：黄睿[1] 李涛[2] 黄敏珍[1] 张敏芳[2] 庄广伦[2] 

机构地区：[1]广东省妇幼保健院生殖医学中心,广州510010 [2]中山大学附属第一医院生殖中心,广州510089

出　　处：《华中科技大学学报（医学版）》2007年第2期220-223,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：广东省自然科学基金资助项目(No.04300799)

摘　　要：目的检测人卵和胚胎中线粒体基因组的表达,探讨线粒体基因表达水平与卵母细胞发育潜能的关系。方法收集临床常规体外授精(IVF)和单精子卵胞浆内注射(ICSI)不受精的成熟卵母细胞以及不适合移植和冷冻保存的胚胎,单个细胞裂解液制备单个卵母细胞或胚胎总RNA,半定量逆转录-多聚酶链式反应(RT-PCR)检测线粒体DNA(mtDNA)编码的5个呼吸链蛋白基因的mRNA含量。结果在所有未受精人卵和胚胎中都检测到了线粒体5种基因的mRNA;未受精人卵ND2、ND5、COⅡ、Cytb、ATP酶6的mRNA含量分别为:(0.38±0.16)、(0.75±0.31)、(0.49±0.22)、(0.74±0.30)(、0.53±0.24),明显低于胚胎中相应5种基因mRNA的含量,其分别为(2.16±0.41)、(3.40±0.53)(、2.40±0.40)(、3.26±0.46)(、2.32±0.42),与胚胎的mRNA含量相比,均P<0.01;三原核胚胎和发育阻滞胚胎的5种基因mRNA水平差异没有显著性意义。结论线粒体基因表达下降可能参与卵母细胞不受精的机制,并与卵母细胞的发育潜能有关。Objective To detect the gene expression level of mitochondrial genome in single human oocyte or embryo, and to explore the relationship between the gene expression level of mtDNA and oocyte developmental potential. Methods Human unfertilized oocytes and embryos unsuitable to transfer were collected. RNA was isolated through cell lyses. Single cell RT-PCR was performed to amplify 5 genes encoding the electric chain subunits. Results These 5 genes＇ mRNAs were detected in all unfertilized oocytes and embryos. The mRNA levels of ND2, ND5, CO Ⅱ , Cytb and ATPase 6 in oocytes were （0.38±0.16）, （0.75±0.31）, （0.49±0.22）, （0.74±0.30）, （0.53±0.24） respectively, significantly lower than those of 5 genes in embryos （ 2.16±0.41, 3.40±0.53, 2.40±0.40, 3.26±0.46, 2.32±0.42, respectively）. There was no significant difference between 3PN embryos and arrested embryos. Conclusion The abnormal expression of mtDNA may involve in the mechanism of fertilization failure and there is some correlation between mtDNA gene expression and oocyte developmental potential.

关 键 词：卵母细胞 发育潜能 DNA 线粒体 基因表达 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]