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Effects of Iloprost on Invitro Development of Preimplantation Porcine Embryos

Author: WangFeng
Tutor: LiuZhongHua
School: Northeast Agricultural University
Course: Developmental Biology
Keywords: iloprost Embryos in Lipid metabolism Pig
CLC: S828
Type: Master's thesis
Year: 2011
Downloads: 29
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Abstract


Cloned pigs with various strains have been born in transgenic cloning technology as an important means of Pig Breeding increasingly wide range of applications. Pig embryo in vitro culture system is still not perfect, a significantly lower proportion of in vitro development of blastocyst and quality, the imperfect embryos in vitro culture system is the production of cloned transgenic pigs inefficient, restrict pig in vitro embryos application one of the factors. Add body factors exogenous to improve in vitro embryo culture system is expected to improve pig embryo development in vitro mass culture system. PGI2 is prostaglandin family a present in the uterine environment in the fallopian tube, play an important role in early embryonic development. In this study, a PGI2 analogue iloprost, explore PGI2 pig embryonic development. Experimental in vitro fertilization (IVF) embryos for research object, respectively, at different times (0h, 24h, 48h, 72h) at different concentrations (0μM, 0.2 μM, 0.5μM, 1 μM, 2 μM 5μM, 10 μM) iloprost added to pig vitro embryo culture medium, blastocyst rates and blastocyst cell number recorded at 156h, screening the best add methods, detection the embryonic fat metabolic rate and metabolism-related genes (cox2, creb, pparδ, pdk, cpt2) expression levels, resolve iloprost affect the mechanism of embryonic development. Experiments to obtain the following results 1) were added at different times 1μM concentration the iloprost, results showed that 48 hours after fertilization, blastocyst rate and blastocyst cell number was significantly greater than the other groups, adding iloprost. 2) at 48 hours after fertilization, respectively, added to different concentrations of iloprost, results showed that the addition of 2μM iloprost blastocyst rate and blastocyst cell number was significantly greater than the other groups. The iloprost Add 48 h after fertilization join 2μM, the the embryonic blastocyst rate (28%) and blastocyst cell number (49.42) was significantly (P lt; 0.05) higher than the rate of blastocyst (16 %) and blastocyst cell number (28.22) 3) added 2μM concentration in culture 48 hours after iloprost, results showed that parthenogenetic embryos, IVF embryos, nuclear transfer embryos between developmental improvement there is no significant difference. 4) application of oil red O staining, detection PGI2 embryo fatty acid content affect, results show add iloprost embryonic fatty acid degradation speed (experimental group was 6.63 × 104vs control group, 8.96 × 104) is also significant with (p-lt; 0.05) to speed up. 5) detection added the iloprost express the amount of energy metabolism genes (cox2, creb, pparδ, pdk, cpt2), the results show a significant increase in the expression levels of fatty acid metabolism-related genes (cox2, creb, pparδ, pdk), glucose metabolism no significant changes in the expression levels of the gene (cpt2). The results showed that the embryos PGI2 by promoting the degradation of fatty acids to provide energy for embryonic development and significantly improve embryo developmental competence.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Pig
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