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Quantitative analysis of mitochondrial DNAs in macaque embryos reprogrammed by rabbit oocytes

State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 25 Bei Si Huan Xi Lu, Haidian, Beijing 100080, China and ¹ Center for Developmental Biology, Shanghai Second Medical University/Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, 1665 Kong Jiang Road, Shanghai 200092, China

Correspondence should be addressed to D-Y Chen; Email: chendy{at}panda.ioz.ac.cn

In cloned animals where somatic cell nuclei and oocytes are from the same or closely related species, the mitochondrial DNA (mtDNA) of the oocyte is dominantly inherited. However, in nuclear transfer (NT) embryos where nuclear donor and oocyte are from two distantly related species, the distribution of the mtDNA species is not known. Here we determined the levels of macaque and rabbit mtDNAs in macaque embryos reprogrammed by rabbit oocytes. Quantification using a real-time PCR method showed that both macaque and rabbit mtDNAs coexist in NT embryos at all preimplantation stages, with maternal mtDNA being dominant. Single NT embryos at the 1-cell stage immediately after fusion contained 2.6 x 10⁴ copies of macaque mtDNA and 1.3 x 10⁶ copies of rabbit mtDNA. Copy numbers of both mtDNA species did not change significantly from the 1-cell to the morula stages. In the single blastocyst, however, the number of rabbit mtDNA increased dramatically while macaque mtDNA decreased. The ratio of nuclear donor mtDNA to oocyte mtDNA dropped sharply from 2% at the 1-cell stage to 0.011% at the blastocyst stage. These results suggest that maternal mtDNA replicates after the morula stage.

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