| As a natural endogenous ligand for G protein-coupled receptors,Ghrelin is widely distributed in tissues and organs of animals with a lot of functions.It also exists in sheep’s ovaries.The existing research results confirm that Ghrelin is expressed in animal oocytes,granulosa cells,and luteal cells.And Ghrelin plays an important role in the development of oocytes and early embryos.In this experiment,we used sheep oocytes as experimental materials to explore the cell cycle regulation effect of Ghrelin-mediated oocyte maturation at different stages of in vitro maturation and the related mechanism of promoting oocyte maturation and the effect of Ghrelin on early embryo development were studied.The results as follow:Add 200ng/mL Ghrelin,which has an effect of improving the in vitro maturation and maturation rate of sheep oocytes(P<0.01)and accelerating the in vitro maturation time(P<0.01).Ghrelin can promote nuclear maturation of sheep oocytes.The transcriptomic analysis showed that Ghrelin promoted the role of Ghrelin by mediating the differential expression of ERK1,ERK2,CCNB2,and CCNB3 signaling pathways in the cell cycle and oocyte meiosis(P<0.05).After in vitro maturation of sheep oocytes obtained after addition of ghrelin,embryonic development after fertilization showed: Ghrelin did not change the order of early embryonic development.Ghrelin reduced the cleavage rate of early embryos(P<0.05).By verifying GLUT1 expression of developmentally related genes,it showed that Ghrelin mediates of GLUT1 expression is declined.(P<0.05).Conclusion:The Ghrelin can accelerate the oocyte culture and maturation in vitro,and through the mediation of ERK1,ERK2,CCNB2,and CCNB3 through PI3K-Akt+MAPK compound signal pathway,Cell Cycle + MAPK compound signal pathway implementation;Ghrelin can reduce early embryo quality and further developmental ability.It was predicted that Ghrelin was down-regulated by GLUT1 gene in the early embryo development process. |
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