Department of Obstetrics and Gynecology, Keio University School of Medicine, 35, Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan,
J Assist Reprod Genet. 2013 Oct;30(10):1367-75. doi: 10.1007/s10815-013-0062-6. Epub 2013 Jul 30.
To investigate the correlation between the ooplasmic volume and the number of mitochondrial DNA (mtDNA) copies in embryos and how they may affect fecundity.
Using real-time PCR, mtDNA quantification was analyzed in unfertilized oocytes and uncleaved embryos. The size of the ovum was also assessed by calculating the ooplasmic volume at the time of granulosa cell removal for IVF or ICSI. Quantification analysis of the mtDNA in blastomeres was performed by real-time PCR at the 7-8 cell stage of the cleaved embryos at 72 h after oocyte retrieval. We calculated the cytoplasmic volume of the blastomeres.
Our studies showed a significantly lower mtDNA copy number in unfertilized oocytes and uncleaved embryos in women who were older than 40 years of age (p < 0.05). The larger ooplasmic volume was also associated with earlier and more rapid cleavage (p < 0.05). The ooplasmic volume was also significantly larger in the group achieving pregnancy. We found a significant positive correlation between blastomere volume and the number of mtDNA copies (r = 0.76, p < 0.01, from Pearson product-moment correlation coefficient).
We have shown that blastomere volume is directly proportional to the number of mtDNA copies. Therefore, larger cytoplasmic volume, with earlier cleavage speed, implies more mtDNA copies. Evaluation of mtDNA quantification and the measurement of ooplasmic and blastomere volume may be useful for selection of high quality embryo and pregnancy outcome.
研究胚胎卵浆体积与线粒体 DNA(mtDNA)拷贝数之间的相关性及其对生育能力的影响。
采用实时 PCR 技术,分析未受精卵和未分裂胚胎中的 mtDNA 定量。在体外受精或卵胞浆内单精子注射时,通过计算颗粒细胞去除时的卵浆体积来评估卵的大小。在卵母细胞回收后 72 小时的分裂胚胎 7-8 细胞期,通过实时 PCR 对分裂胚胎的卵裂球 mtDNA 进行定量分析。我们计算了卵裂球的细胞质体积。
我们的研究表明,年龄大于 40 岁的女性中,未受精卵和未分裂胚胎的 mtDNA 拷贝数明显较低(p<0.05)。较大的卵浆体积也与更早和更快的卵裂有关(p<0.05)。妊娠组的卵浆体积也明显更大。我们发现卵裂球体积与 mtDNA 拷贝数之间存在显著的正相关关系(r=0.76,p<0.01,皮尔逊积矩相关系数)。
我们已经表明,卵裂球体积与 mtDNA 拷贝数成正比。因此,较大的细胞质体积和较快的卵裂速度意味着更多的 mtDNA 拷贝数。评估 mtDNA 定量和卵浆及卵裂球体积的测量可能有助于选择高质量的胚胎和妊娠结局。