Shitara H, Kaneda H, Sato A, Inoue K, Ogura A, Yonekawa H, Hayashi J I
Department of Laboratory Animal Science, Tokyo Metropolitan Institute of Medical Science, Tokyo 113-8613, Japan.
Genetics. 2000 Nov;156(3):1277-84. doi: 10.1093/genetics/156.3.1277.
Exclusion of paternal mitochondria in fertilized mammalian eggs is very stringent and ensures strictly maternal mtDNA inheritance. In this study, to examine whether elimination was specific to sperm mitochondria, we microinjected spermatid or liver mitochondria into mouse embryos. Congenic B6-mt(spr) strain mice, which are different from C57BL/6J (B6) strain mice (Mus musculus domesticus) only in possessing M. spretus mtDNA, were used as mitochondrial donors. B6-mt(spr) mice and a quantitative PCR method enabled selective estimation of the amount of M. spretus mtDNA introduced even in the presence of host M. m. domesticus mtDNA and monitoring subsequent changes of its amount during embryogenesis. Results showed that M. spretus mtDNA in spermatid mitochondria was not eliminated by the blastocyst stage, probably due to the introduction of a larger amount of spermatid mtDNA than of sperm mtDNA into embryos on fertilization. However, spermatid-derived M. spretus mtDNA was eliminated by the time of birth, whereas liver-derived M. spretus mtDNA was still present in most newborn mice, even though its amount introduced was significantly less than that of spermatid mtDNA. These observations suggest that mitochondria from spermatids but not from liver have specific factors that ensure their selective elimination and resultant elimination of mtDNA in them, and that the occurrence of elimination is not limited to early stage embryos, but continues throughout embryogenesis.
在受精的哺乳动物卵子中排除父系线粒体非常严格,可确保严格的母系线粒体DNA遗传。在本研究中,为了检验这种排除是否对精子线粒体具有特异性,我们将精子细胞或肝脏线粒体显微注射到小鼠胚胎中。同基因B6-mt(spr)品系小鼠仅在拥有西班牙小家鼠线粒体DNA方面与C57BL/6J(B6)品系小鼠(小家鼠)不同,被用作线粒体供体。B6-mt(spr)小鼠和定量PCR方法能够选择性地估计即使在存在宿主小家鼠线粒体DNA的情况下引入的西班牙小家鼠线粒体DNA的量,并监测其在胚胎发育过程中随后的量的变化。结果表明,精子细胞线粒体中的西班牙小家鼠线粒体DNA在囊胚阶段并未被排除,这可能是由于受精时引入胚胎的精子细胞线粒体DNA的量比精子线粒体DNA的量更多。然而,精子细胞来源的西班牙小家鼠线粒体DNA在出生时被排除,而肝脏来源的西班牙小家鼠线粒体DNA在大多数新生小鼠中仍然存在,尽管其引入量明显少于精子细胞线粒体DNA。这些观察结果表明,精子细胞而非肝脏的线粒体具有特定因子,可确保它们被选择性排除并导致其中的线粒体DNA被排除,并且排除的发生不限于早期胚胎,而是在整个胚胎发育过程中持续存在。