Anti-apoptotic effect of melatonin on preimplantation development of porcine parthenogenetic embryos

被引:88
作者
Choi, Jiho [1 ]
Park, Seon-Mi [1 ]
Lee, Eugine [1 ]
Kim, Ji-Hye [1 ]
Jeong, Yeon-Ik [1 ]
Lee, Jong-Yun [1 ]
Park, Sun-Woo [1 ]
Kim, Huen-Suk [1 ]
Hossein, Mohammad Shamim [1 ]
Jeong, Yeon-Woo [1 ]
Kim, Sue [1 ]
Hyun, Sang-Hwan [1 ,2 ]
Hwang, Woo-Suk [1 ]
机构
[1] SooAm Biotech Res Fdn, Seoul 137851, South Korea
[2] Chungbuk Natl Univ, Coll Vet Med, Lab Vet Biotechnol, Cheongju, South Korea
关键词
parthenogenetic development; somatic cell nuclear transfer; melatonin; reactive oxygen species; apoptosis;
D O I
10.1002/mrd.20861
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the present study, we investigated the effect of melatonin on the preimplantation development of porcine parthenogenetic and somatic cell nuclear transfer (SCNT) embryos. Parthenogenetic embryos were cultured in mNCSU-23 supplemented with various concentrations of melatonin for 7 days. The results revealed that 100 pM was the optimal concentration, which resulted in significantly increased cleavage and blastocyst formation rates. Additionally, 100 pM melatonin provided the highest increase in total cell number of blastocysts. Therefore, the subsequent experiments were performed with 100 pM melatonin. ROS level in 2-8 cell stage embryos in the presence or absence of melatonin was evaluated. Embryos cultured with melatonin showed significantly decreased ROS. Blastocysts cultured with melatonin for 7 days were analyzed by the TUNEL assay. It was observed that melatonin not only increased (P < 0.05) the total cell number but also decreased (P<0.05) the rate of apoptotic nuclei. Blastocysts cultured with melatonin were assessed for the expression of apoptosis-related genes Bcl-xl and Bax, and of pluripotency marker gene Oct-4 by real-time quantitative PCR. Analysis of data showed that the expression of Bcl-xl was higher (1.7-fold) compared to the control while the expression of Bax was significantly decreased relative to the control (0.7-fold) (P < 0.05). Moreover, the expression of Oct-4 was 1.7-fold higher than the control. These results indicated that melatonin had beneficial effects on the development of porcine parthenogenetic embryos. Based on the findings of parthenogenetic embryos, we investigated the effect of melatonin on the development of porcine SCNT embryos. The results also demonstrated increased cleavage and blastocyst formation rates, and the total cell numbers in blastocysts were significantly higher when the embryos were cultured with melatonin. Therefore, these data suggested that melatonin may have important implications for improving porcine preimplantation SCNT embryo development.
引用
收藏
页码:1127 / 1135
页数:9
相关论文
共 59 条
[1]   The Bcl-2 protein family: Arbiters of cell survival [J].
Adams, JM ;
Cory, S .
SCIENCE, 1998, 281 (5381) :1322-1326
[2]   Neurohormone melatonin prevents cell damage: Effect on gene expression for antioxidant enzymes [J].
Antolin, I ;
Rodriguez, C ;
Sainz, RM ;
Mayo, JC ;
Uria, H ;
Kotler, MI ;
RodriguezColunga, MJ ;
Tolivia, D ;
MenendezPelaez, A .
FASEB JOURNAL, 1996, 10 (08) :882-890
[3]   Inhibition of Bax channel-forming activity by Bcl-2 [J].
Antonsson, B ;
Conti, F ;
Ciavatta, A ;
Montessuit, S ;
Lewis, S ;
Martinou, I ;
Bernasconi, L ;
Bernard, A ;
Mermod, JJ ;
Mazzei, G ;
Maundrell, K ;
Gambale, F ;
Sadoul, R ;
Martinou, JC .
SCIENCE, 1997, 277 (5324) :370-372
[4]   Oct4 distribution and level in mouse clones:: consequences for pluripotency [J].
Boiani, M ;
Eckardt, S ;
Schöler, HR ;
McLaughlin, KJ .
GENES & DEVELOPMENT, 2002, 16 (10) :1209-1219
[5]   BCL-X, A BCL-2-RELATED GENE THAT FUNCTIONS AS A DOMINANT REGULATOR OF APOPTOTIC CELL-DEATH [J].
BOISE, LH ;
GONZALEZGARCIA, M ;
POSTEMA, CE ;
DING, LY ;
LINDSTEN, T ;
TURKA, LA ;
MAO, XH ;
NUNEZ, G ;
THOMPSON, CB .
CELL, 1993, 74 (04) :597-608
[6]  
Booth P J, 2001, Cloning Stem Cells, V3, P139, DOI 10.1089/153623001753205098
[7]   Calcium, ATP, and ROS: a mitochondrial love-hate triangle [J].
Brookes, PS ;
Yoon, YS ;
Robotham, JL ;
Anders, MW ;
Sheu, SS .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2004, 287 (04) :C817-C833
[8]   Apaf1 (CED-4 homolog) regulates programmed cell death in mammalian development [J].
Cecconi, F ;
Alvarez-Bolado, G ;
Meyer, BI ;
Roth, KA ;
Gruss, P .
CELL, 1998, 94 (06) :727-737
[9]   Effect of different parthenogenetic activation methods on the developmental competence of in vitro matured porcine oocytes [J].
Cheng, Wen-Min ;
Sun, Xu-Lei ;
An, Lei ;
Zhu, Shi-En ;
Li, Xi-He ;
Li, Ying ;
Tian, Jian-Hui .
ANIMAL BIOTECHNOLOGY, 2007, 18 (02) :131-141
[10]  
Colman A, 1999, Cloning, V1, P185