Sphingoshine-1-Phosphate Enhances Meiotic Maturation and Further Embryonic Development in Pigs
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서지정보
ㆍ발행기관 : 한국동물번식학회
ㆍ수록지정보 : Reproductive & developmental biology / 36권 / 3호
ㆍ저자명 : Hyo-Sang Lee, Deog-Bon Koo
ㆍ저자명 : Hyo-Sang Lee, Deog-Bon Koo
목차
ABSTRACT INTRODUCTION MATERIALS AND METHODS Treatment of SlP and DMS In Vitro Maturation (IVM) In Vitro Fertilization (IVF) and Culture (IVC) Aceto-Orcein Staining Western Blot Analysis TUNEL and Cell Counting Statistical Analysis RESULTS Effect of S1P Treatment during IVM of Porcine Oocytes on Meiotic Maturation and Embryo Development S1P Induces MAP Kinase Phosphorylation in In Vitro Matured Porcine Oocytes Effect of S1P during IVC on Embryo Development and Apoptosis Effect of S1P during IVM and IVC Periods on Embryo Development DISCUSSION REFERENCES영어 초록
Sphingosine-1-phosphate (S1P) has a many function involved proliferation, differentiation and survival of many cells. In this study, to investigate whether S1P improve the developmental competence of porcine embryos, 50 nM of S1P were supplemented during in vitro maturation (with EGF or without EGF) medium and/or in vitro culture (IVC) medium. Addition of S1P was significantly increased the rate of oocytes reaching metaphase II (MII) compared to the control (83.5 vs. 64.1%) in without EGF medium, but not with EGF medium (89.5 vs. 84.6%). When treated with 1 μM of N1N-dimethylsphingosine (DMS), a sphingosine kinase inhibitor which is blocked endogenous generation of S1P, the meiotic progression rates to MII stage (without EGF: 45.2 and with EGF: 66.7%) were significantly decreased and degeneration rates (without EGF: 51.2 and with EGF: 30.1%) were increased in both medium compared to control group during IVM periods. Also, the rates of blastocyst formation was significantly increased in the S1P treated group compared to control group (29.0 vs. 19.2%) of EGF supplemented medium, whereas there were no effect in the EGF free medium (9.0 vs. 10.5%). After 12 h IVM, the phosphorylation of ERK1 and ERK2, which is major signaling pathway of MAP kinase, were increased in the S1P group than that of control or DMS group. When supplemented of S1P during IVC, the rates of blastocyst formation and total cell number (30.2% and 40.6) were significantly increased in S1P-treated group compared with control (20.1% and 32.5), DMS (12.3% and 25.1), and S1P plus DMS group (24.7% and 33.6). The percentage of apoptosis nuclei in the S1P group was significantly decreased than other groups. Also, the rates of blastocyst formation (26.7 vs. 14%) and total cell number (42.8 vs. 32.5) were significantly increased in the S1P group than those of control group when S1P added during the entire IVM and IVC periods. Taken together, our results indicate that S1P supplementation in IVM and/or IVC medium affects beneficial effect of meiotic maturation and subsequent developmental competence of porcine embryos.참고 자료
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