Mitochondria directly influence fertilisation outcome in the pig
Mitochondria directly influence fertilisation outcome in the pig
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DOI：
10.1530/rep.1.00551
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发表时间：
2006-02-01
期刊：
影响因子：
3.8
通讯作者：
St John, JC
中科院分区：
文献类型：
--
作者：
El Shourbagy, SH;Spikings, EC;St John, JC
它明确参与细胞质调节，是细胞ATP的主要产生者。我们的目的是确定线粒体是否单独影响受精结果。在体外，如染料亮甲酚蓝（BCB）所示，卵母细胞的能力可以通过葡萄糖-6-磷酸脱氢酶（G6 PD）的存在来评估。使用猪体外受精（IVF），我们评估了卵母细胞成熟，细胞质体积，受精结果，线粒体数量确定的mtDNA拷贝数，以及线粒体是否均匀分布在每个胚胎的卵裂球。在用BCB染色后，我们观察到BCB阳性（BCB+）和RCB阴性（BCB-）卵母细胞的胞质体积存在显著差异。受精卵和未受精卵之间的mtDNA拷贝数也有显着差异，卵裂早期卵裂球之间的线粒体分离不平等。此外，我们用来自母系亲属的线粒体补充BCB-卵母细胞，并观察到补充的、假注射的和未处理的BCB-卵母细胞之间在IVF和胞浆内单精子注射（ICSI）后的受精结果的显著差异。因此，我们已经证明了卵母细胞成熟度，细胞质体积，受精结果和线粒体含量之间的关系。这些数据表明，线粒体数量对受精结果和胚胎发育很重要。此外，线粒体受精前阈值可以确保，由于线粒体在受精后卵裂期间被稀释，每个卵裂球有足够的mtDNA拷贝，以允许在早期植入后阶段期间启动mtDNA复制后将mtDNA传输到植入后胚胎的每个细胞。
The mitochondrion is explicitly involved in cytoplasmic regulation and is the cell's major generator of ATP. Our aim was to determine whether mitochondria alone could influence fertilisation outcome. in vitro, oocyte competence can be assessed through the presence of glucose-6-phosphate dehydrogenase (G6PD) as indicated by the dye, brilliant cresyl blue (BCB). Using porcine in vitro fertilisation (IVF), we have assessed oocyte maturation, cytoplasmic volume, fertilisation outcome, mitochondrial number as determined by mtDNA copy number, and whether mitochondria are uniformly distributed between blastomeres of each embryo. After staining with BCB, we observed a significant difference in cytoplasmic volume between BCB positive (BCB+) and RCB negative (BCB-) oocytes. There was also a significant difference in mtDNA copy number between fertilised and unfertilised oocytes and unequal mitochondrial segregation between blastomeres during early cleavage stages. Furthermore, we have supplemented BCB- oocytes with mitochondria from maternal relatives and observed a significant difference in fertilisation outcomes following both IVF and intracytoplasmic sperm injection (ICSI) between supplemented, sham-injected and non-treated BCB- oocytes. We have therefore demonstrated a relationship between oocyte maturity, cytoplasmic volume, and fertilisation outcome and mitochondrial content. These data suggest that mitochondrial number is important for fertilisation outcome and embryonic development. Furthermore, a mitochondrial pre-fertilisation threshold may ensure that, as mitochondria are diluted out during post-fertilisation cleavage, there are sufficient copies of mtDNA per blastomere to allow transmission of mtDNA to each cell of the post-implantation embryo after the initiation of mtDNA replication during the early postimplantation stages.