Original Article

Mito-TEMPO에 의한 미토콘드리아 유래 초과산화물의 감소가 돼지 난모세포 성숙에 미치는 영향


Reduction of Mitochondrial Derived Superoxide by Mito-TEMPO Improves Porcine Oocyte Maturation In Vitro

양슬기, 박효진, 이상민, 김진우, 김민지, 김인수, 제갈호근, 구덕본

피인용수 : 0(자료제공 : 네이버학술정보)



Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: 79.9 ± 3.8% vs G2: 57.5 ± 4.6%) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO (0.1 μM, MT) treatment (G2: 68.4 ± 3.2% vs G2 + MT: 73.9 ± 1.4%). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.


재료 및 방법
난소 채취 및 난포란 체외성숙
난구세포 정량
난포란의 핵 성숙 확인
Mito-SOX 염색
RNA 추출 및 역전사 PCR (RT-PCR)
통계 분석
난구세포의 수에 따른 돼지 난포란의 핵 성숙율 확인
성숙된 난포란에서 초과산화물 생성과 항산화 효소의 발현 조사
Mito-TEMPO 첨가에 따른 돼지 난포란의 핵 성숙율 변화 확인


  • 양슬기 Seul-Gi Yang. 대구대학교 공과대학 생명공학과
  • 박효진 Hyo-Jin Park. 대구대학교 공과대학 생명공학과
  • 이상민 Sang-Min Lee. 대구대학교 공과대학 생명공학과
  • 김진우 Jin-Woo Kim. 대구대학교 공과대학 생명공학과
  • 김민지 Min-Ji Kim. 대구대학교 공과대학 생명공학과
  • 김인수 In-Su Kim. 대구대학교 공과대학 생명공학과
  • 제갈호근 Ho-Geun Jegal. 대구대학교 공과대학 생명공학과
  • 구덕본 Deog-Bon Koo. 대구대학교 공과대학 생명공학과


자료제공 : 네이버학술정보

    함께 이용한 논문

      ※ 기관로그인 시 무료 이용이 가능합니다.

      • 4,000원

      0개의 논문이 장바구니에 담겼습니다.