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Reproductive & Developmental Biology(Supplement)

간행물 정보
  • 자료유형
    학술대회
  • 발행기관
    한국동물생명공학회(구 한국동물번식학회) [The Korean Society of Animal Reproduction and Biotechnology]
  • 간기
    연간
  • 수록기간
    2001 ~ 2017
  • 주제분류
    농수해양 > 축산학
  • 십진분류
    KDC 527 DDC 636
Volume 38 No 2 Supplement (114건)
No
72

Cathepsin B, a lysosomal cysteine protease of the papain family, has recently been implicated in quality and developmental competence of bovine preimplantation embryos. In this study, to determine whether inhibition of Cathepsin B activity could improve oocyte maturation and early embryo developmental competence in porcine, we added E-64, an inhibitor of Cathepsin B, into the in vitro maturation and culture media. Both mRNA and protein expression were no differences in germinal vesicle (GV) stage oocytes between good and poor quality from the morphology. Even activity of Cathepsin B is high in good quality GV stage oocytes, E-64 did not enhance in vitro maturation of porcine oocyte. However, I nM E-64 significantly (p<0.05) increased blastocyst formation and cell numbers. Inhibition of Cathepsin B activity did not alter mitochondrial membrane potential, copy numbers and ATP contents. But, E-64 significantly (p<0.05) reduced mitochondrial release of cytochrome c, finally decreased mRNA expression of Caspase 3 and Caspase 9, resulted in reduction of apoptotic index in porcine blastocysts. Collectively, these observations suggest that Inhibition of Cathepsin B activity enhanced developmental competence and reduced apoptosis through protecting against cytochrome C release from mitochondria in porcine parthenotes.

73

Iron is an important element which takes part in several biological processes. The role of iron in porcine oocyte maturation and early embryo development was still unknown. In the present study, iron was depleted by deferoxamine (DFM), a specific iron chelator, during in vitro maturation (IVM) and in vitro culture (IVC) period respectively. Results showed that, oocytes were matured with 1 μM DFM for 44 h followed by parthenogenetic activation (PA) displayed higher (p<0.05) blastocyst rate compared to without DFM in IVM medium. Another hand, supplemented with 0.5 μM DFM to the IVC medium for 7 days enhanced blastocyst formation both in PA and in vitro fertilization (IVF) groups (p<0.05). Although there was no difference in total cell number in IVF-derived blastocysts between control and iron depleted group, apoptotic index was significantly (p<0.05) decreased following iron depletion. ROS content in MII oocyte and blastocyst was significantly (p<0.05) decreased after iron depletion. For clarify the mechanism of the phenomenon, expressions of apoptotic- and ROS-related genes in the blastocyst were checked. Iron depletion reduced the expression of Caspase 3 and increased Bcl-xL in blastocyst (p<0.05), but there was no significanly differenc in m-RNA expression of Nox4. Mitochondrial membrane potential was increased both in MII oocytes and blastocysts after DFM treatment. In conclusion, iron depletion decreased ROS in oocytes and embryos developing in vitro, further enhanced mitochondrial activity and reduced the apoptosis in blastosyst, resulted in enhancement of the developmental capacity in porcine early embryos.

77

Fetal Bovine Serum Improves Developmental Competance of Bovine Early Embryos through Alleviating Oxidative Stress

Seong-Eun Mun, Pil-Soo Jeong, Hae-Jun Yang, Seon-A Choi, Seung-Bin Yoon, Young-Ho Park, Bong-Seok Song, Bo-Woong Sim, Ji-Su Kim, Sun-Uk Kim, Kyu-Tae Chang

한국동물생명공학회(구 한국동물번식학회) Reproductive & Developmental Biology(Supplement) Volume 38 No 2 Supplement 2014.06 p.110

84

Embryonic stem (ES) cells are pluripotent cells with the capacity for unlimited self-renewal or differentiation. Inhibition of MAPK pathways enhances mouse ES cell pluripotency characteristics. Compared to wildtype ES cells, jnk2(—/—) ES cells displayed a much higher growth rate. To determine whether JNKs are required for stem cell selfrenewal or differentiation, we performed a phosphorylation kinase array assay to compare mouse ES cells under LIF+ or LIF- culture conditions. The data showed that activation of JNKs was induced by LIF withdrawal. We also found that JNK1 or 2 phosphorylated Klf4 at threonines 224 and 225. Activation of JNK signaling and phosphorylation of Klf4 inhibited Klf4 transcription and transactivation activity. Importantly, jnk1 (—/—) and jnk2(—/—) murine embryonic fibroblasts (MEFs) exhibited a significantly greater potency in the ability to increase the number of iPS colonies compared with jnkwildtype MEFs. Overall, our results demonstrated that JNK1 and 2 play a negative role in reprogramming to pluripotent stem cells by suppressing Klf4 activity.

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