Mitochondria transfer into mouse ova by microinjection
Mitochondria transfer into mouse ova by microinjection
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DOI：
10.1023/a:1018431316831
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发表时间：
1997-11-01
影响因子：
3
通讯作者：
Moffatt, RJ
中科院分区：
文献类型：
--
作者：
Pinkert, CA;Irwin, MH;Moffatt, RJ
建立了一种小鼠线粒体分离和种间移植的方法。从小家鼠肝脏标本中提取线粒体，显微注射到超数排卵的家蝇雌性受精卵中。用于显微注射的线粒体制剂的电子显微镜观察显示，线粒体囊泡完整，微粒体污染很少。与线粒体DNA D-loop区序列差异互补的物种特异性嵌套PCR引物显示，分离并注射到胚胎发育的囊胚期培养的受精卵后，外源线粒体的成功率很高。体外培养4.5d后，217个受精卵中有67个存活，37个发育的受精卵中有23个处于胚胎发育的囊胚期。所有23个胚泡都含有可检测到的外来线粒体。这些结果是开发研究线粒体动力学的模型系统的第一步，并为受线粒体功能异常或突变影响的人类代谢性疾病开发治疗策略。
A method for mitochondria isolation and interspecific transfer of mitochondria was developed in mice. Mitochondria were isolated from Mus spretus liver samples for microinjection into fertilized ova obtained from superovulated M. musculus domesticus females. Electron microscopic observations of mitochondria preparations used for microinjection demonstrated intact mitochondrial vesicles with little microsomal contamination. Species-specific nested PCR primers complementary to sequence differences in the mitochondrial DNA D-loop region revealed high rates of successful transfer of foreign mitochondria after isolation and injection into zygotes cultured through the blastocyst stage of embryonic development. Of 217 zygotes, 67 survived mitochondria injection and 23 out of 37 zygotes developed were at the blastocyst-stage of embryonic development after 4.5 days of in vitro culture. All 23 of these blastocysts contained detectable levels of foreign mitochondria. These results represent an initial step in developing a model system to study mitochondrial dynamics and development of therapeutic strategies for human metabolic diseases affected by aberrations in mitochondrial function or mutation.