College of Animal Science and Technology, China Agricultural University, 100193 Beijing, China.
Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, 610041 Chengdu, China.
Biol Chem. 2019 Apr 24;400(5):613-623. doi: 10.1515/hsz-2018-0273.
Most animals generated by somatic cell nuclear transfer (SCNT) are heteroplasmic; inheriting mitochondrial genetics from both donor cells and recipient oocytes. However, the mitochondrial genome and functional mitochondrial gene expression in SCNT animals are rarely studied. Here, we report the production of SCNT pigs to study introduction, segregation, persistence and heritability of mitochondrial DNA transfer during the SCNT process. Porcine embryonic fibroblast cells from male and female Xiang pigs were transferred into enucleated oocytes from Yorkshire or Landrace pigs. Ear biopsies and blood samples from SCNT-derived pigs were analyzed to characterize the mitochondrial genome haplotypes and the degree of mtDNA heteroplasmy. Presence of nuclear donor mtDNA was less than 5% or undetectable in ear biopsies and blood samples in the majority of SCNT-derived pigs. Yet, nuclear donor mtDNA abundance in 14 tissues in F0 boars was as high as 95%. Additionally, mtDNA haplotypes influenced mitochondrial respiration capacity in F0 fibroblast cells. Our results indicate that the haplotypes of recipient oocyte mtDNA can influence mitochondrial function. This leads us to hypothesize that subtle developmental influences from SCNT-derived heteroplasmy can be targeted when using donor and recipient mitochondrial populations from breeds of swine with limited evolutionary divergence.
大多数通过体细胞核移植（SCNT）产生的动物都是异质的；它们从供体细胞和受体卵母细胞中继承了线粒体遗传学。然而，SCNT 动物中的线粒体基因组和功能性线粒体基因表达很少被研究。在这里，我们报告了 SCNT 猪的生产，以研究线粒体 DNA 转移在 SCNT 过程中的引入、分离、持续存在和遗传。来自雄性和雌性湘猪的胚胎成纤维细胞被转移到来自约克夏或长白猪的去核卵母细胞中。对 SCNT 衍生猪的耳活检和血液样本进行分析，以表征线粒体基因组单倍型和 mtDNA 异质性程度。在大多数 SCNT 衍生猪的耳活检和血液样本中，核供体 mtDNA 的存在少于 5%或无法检测到。然而，F0 公猪 14 种组织中的核供体 mtDNA 丰度高达 95%。此外，mtDNA 单倍型影响 F0 成纤维细胞的线粒体呼吸能力。我们的结果表明，受体卵母细胞 mtDNA 的单倍型可以影响线粒体功能。这使我们假设，当使用来自进化分歧有限的猪品种的供体和受体线粒体群体时，可以针对 SCNT 衍生异质体的细微发育影响进行靶向。
