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KMID : 0613820170270121445
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Journal of Life Science
2017 Volume.27 No. 12 p.1445 ~ p.1451
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Transfer of Isolated Mitochondria to Bovine Oocytes by Microinjection
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Baek Sang-Ki
Byun June-Ho
Kim Bo-Gyu
Lee A-Ram
Cho Young-Su
Kim Ik-Sung
Seo Gang-Mi
Chung Se-Kyo
Lee Joon-Hee
Woo Dong-Kyun
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Abstract
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Mitochondria play a central role in energy generation by using electron transport coupled with oxidative phosphorylation. They also participate in other important cellular functions including metabolism, apoptosis, signaling, and reactive oxygen species production. Therefore, mitochondrial dysfunction is known to contribute to a variety of human diseases. Furthermore, there are various inherited diseases of energy metabolism due to mitochondrial DNA (mtDNA) mutations. Unfortunately, therapeutic options for these inherited mtDNA diseases are extremely limited. In this regard, mitochondrial replacement techniques are taking on increased importance in developing a clinical approach to inherited mtDNA diseases. In this study, green fluorescence protein (GFP)-tagged mitochondria were isolated by differential centrifugation from a mammalian cell line. Using microinjection technique, the isolated GFP-tagged mitochondria were then transferred to bovine oocytes that were triggered for early development. During the early developmental period from bovine oocytes to blastocysts, the transferred mitochondria were observed using fluorescent microscopy. The microinjected mitochondria were dispersed rapidly into the cytoplasm of oocytes and were passed down to subsequent cells of 2-cell, 4-cell, 8-cell, morula, and blastocyst stages. Together, these results demonstrate a successful in vitro transfer of isolated mitochondria to oocytes and provide a model for mitochondrial replacement implicated in inherited mtDNA diseases and animal cloning.
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KEYWORD
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Development, disease, microinjection, mitochondria, mitochondrial replacement
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