title: Anti-apoptotic effect of melatonin on preimplantation development of porcine parthenogenetic embryos.
abstract: 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.
authors.affiliation: SooAm Biotech Research Foundation, Bangbae3-dong, Seocho-gu, Seoul, Korea.
authors.forename: Jiho
authors.initials: J
authors.lastname: Choi
authors.forename: Seon-Mi
authors.initials: SM
authors.lastname: Park
authors.forename: Eugine
authors.initials: E
authors.lastname: Lee
authors.forename: Ji-Hye
authors.initials: JH
authors.lastname: Kim
authors.forename: Yeon-Ik
authors.initials: YI
authors.lastname: Jeong
authors.forename: Jong-Yun
authors.initials: JY
authors.lastname: Lee
authors.forename: Sun-Woo
authors.initials: SW
authors.lastname: Park
authors.forename: Huen-Suk
authors.initials: HS
authors.lastname: Kim
authors.forename: Mohammad Shamim
authors.initials: MS
authors.lastname: Hossein
authors.forename: Yeon-Woo
authors.initials: YW
authors.lastname: Jeong
authors.forename: Sue
authors.initials: S
authors.lastname: Kim
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
authors.forename: Woo-Suk
authors.initials: WS
authors.lastname: Hwang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D020287:In Situ Nick-End Labeling; D008550:Melatonin / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D010312:Parthenogenesis / Q000187:drug effects / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D013552:Swine
chemical_list: D008550:Melatonin