Songsasen N, Henson L H, Tipkantha W, Thongkittidilok C, Henson J H, Chatdarong K, Comizzoli P
Center for Species Survival, Smithsonian Conservation Biology Institute, National Zoological Park, Front Royal, VA, USA.
Zoological Park Organization, Bangkok, Thailand.
Reprod Domest Anim. 2017 Apr;52 Suppl 2:71-76. doi: 10.1111/rda.12851. Epub 2017 Jan 22.
Mitochondria play fundamental roles during oocyte development. The accumulation and spatial redistribution of these energy-producing organelles have been linked to the developmental competence of mammalian oocytes. Here, we assessed the copy number, distribution and activity of mitochondria within cat oocytes during folliculogenesis. In Experiment 1, oocytes were recovered from primordial (n = 152), primary (112), secondary (95), early (131), small (118), antral (86) and advanced antral (5) stages follicles, and mitochondria DNA extracted and quantified using qPCR. In Experiment 2, oocytes from pre-antral (n = 44), early antral (n = 66), small antral (n = 59), antral (n = 41) and advanced antral (n = 21) follicles were isolated and stained with CMXRos MitoTracker dye to assess mitochondrial distribution pattern and activity levels. Oocyte's mitochondria DNA (mtDNA) copy numbers gradually increased as folliculogenesis progressed, with a significant shift at the small antral stage (0.5 to <1 mm in diameter). The location of mitochondria gradually shifted from a homogeneous distribution throughout the cytoplasm in pre-antral oocytes to a pericortical concentration in the advanced antral stage. Quantification of CMXRos fluorescent intensity revealed a progressive increase in mitochondrial activity in oocytes from the pre-antral to the large antral follicles. Taken together, these findings demonstrated that cat oocytes undergo dynamic changes in mitochondrial copy number, distribution and activity during folliculogenesis. These significant modifications to this crucial cytoplasmic organelle are likely associated with the acquisition of developmental competency by cat oocytes.
线粒体在卵母细胞发育过程中发挥着重要作用。这些产生能量的细胞器的积累和空间重新分布与哺乳动物卵母细胞的发育能力有关。在此,我们评估了猫卵泡发生过程中卵母细胞内线粒体的拷贝数、分布和活性。在实验1中,从原始卵泡(n = 152)、初级卵泡(112个)、次级卵泡(95个)、早期卵泡(131个)、小卵泡(118个)、窦状卵泡(86个)和高级窦状卵泡(5个)中回收卵母细胞,并使用qPCR提取和定量线粒体DNA。在实验2中,分离出前窦状卵泡(n = 44)、早期窦状卵泡(n = 66)、小窦状卵泡(n = 59)、窦状卵泡(n = 41)和高级窦状卵泡(n = 21)中的卵母细胞,并用CMXRos线粒体追踪染料染色,以评估线粒体的分布模式和活性水平。随着卵泡发生的进展,卵母细胞的线粒体DNA(mtDNA)拷贝数逐渐增加,在小窦状卵泡阶段(直径0.5至<1毫米)有显著变化。线粒体的位置从前窦状卵泡卵母细胞中整个细胞质的均匀分布逐渐转移到高级窦状卵泡阶段的皮质周围集中分布。CMXRos荧光强度的定量分析显示,从前窦状卵泡到大型窦状卵泡的卵母细胞中,线粒体活性逐渐增加。综上所述,这些发现表明,猫卵母细胞在卵泡发生过程中线粒体的拷贝数、分布和活性会发生动态变化。对这个关键细胞质细胞器的这些重大改变可能与猫卵母细胞发育能力的获得有关。