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Biology of reproduction, v.74 no.3, 2006년, pp.601 - 610
Cui, Xiang-Shun (National Research Laboratory, Department of Animal Sciences,Chungbuk National University, Chungbuk 361？?763, South Korea) , Li, Xing-Yu (Laboratory of Reproductive Biology & Infertility, Samsung CheilHospital & Women's Healthcare Center, Sungkyunkwan UniversitySchool of Medicine, Seoul 100？?380, Korea) , Jeong, Yu-Jeong , Jun, Jin-Hyun , Kim,Nam-Hyung
To investigate the role of nuclear encoded genes in mitochondrialfunction during oocyte maturation and early embryogenesis weexamined the expression pattern and function of the cytochromeoxidase (Cox) subunits, Cox5a, 5b, and 6b1 during oocytematuration and early embryo development. Transcription of Cox5a,5b, or 6b1 was observed in oocytes and during early development;their expression levels were abundant in mature oocytes (MII) andzygotes (1C), and lowest at the 2-cell stage (2C), graduallyincreasing from 4-cell to blastocyst stage. Immunocytochemicalstudies revealed that COX5A, 5B, or 6B1 proteins were expressed inall blastomeres of the blastocyst. Silencing of mRNA expression byRNA interference (siRNA) did not inhibit oocyte maturation ordevelopmental events up to the morula and blastocyst stages, butdisrupted mitochondrial distribution. Significantly higherapoptosis and lower cell numbers were observed in siRNA-treatedblastocysts. Real time RT-PCR revealed that silencing of Cox5a,5b, or 6b1 did not alter mRNA levels of Bcl-xL (Bcl2l1), butincreased transcription levels of proapoptotic genes, Bax andcaspase 3 (Casp3). Furthermore, mRNA and protein levels ofE-cadherin (CDH1) were decreased in siRNA microinjectedblastocysts. These results suggest that gene expression of the Coxsubunits, Cox5a, 5b, and 6b1 is not required for embryodevelopmental events up to the blastocyst stage. The loss of thesegenes leads to mitochondrial dysfunction that results in apoptosisof the blastocyst stage embryos.
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