ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2015, Vol. 46 ›› Issue (10): 1766-1774.doi: 10.11843/j.issn.0366-6964.2015.10.009

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Function Analysis of Buffalo BMP1 Gene on Proliferation and Apoptosis of Granulosa Cells

LEI Xiao-can,ZHANG Hai-hang,CUI Kui-qing,LIU Fu-hang,LIU Qing-you*,SHI De-shun*   

  1. (State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources,Guangxi University,Nanning 530005,China)
  • Received:2015-02-05 Online:2015-10-23 Published:2015-10-23

Abstract:

This study focus on the affection of BMP1 gene on proliferation and apoptosis of in vitro cultured buffalo granulosa cells.Expression pattern of BMP1 in different in vitro cultured time of buffalo granulosa cells including 0,24,48,72 and 96 h were determined by qRT-PCR separately.Different concentrations of BMP1 recombinant protein or BMP1 antibody were added to the culture medium,and the effect to proliferation of granulose cells were detected,further effect on expression pattern of cell proliferation and apoptosis associated genes were also analyzed by qRT-PCR and Western blotting.BMP1 gene was significantly increased in the in vitro cultured granulosa cells with the extended recording time(P<0.05),and reached a peak at 96 h.Adding BMP1 recombinant protein significantly promoted the proliferation of granulosa cells(P<0.05),as well as up-regulated the expression of Cyclin D1,Cyclin D2 and PCNA genes in mRNA level.In addition,mRNA expression level of pro-apoptosis factors(FasFasLTNFR1, TNF-α,Cytochrome C,Apaf1,ChopCaspase-3,Caspase-8 and Caspase-9) were down regulated.Furthermore,Western blotting results showed that the protein expression level of FasBax and Caspase-9 was also down-regulated.The contrast results were observed by adding BMP1 antibody investigation.BMP1 promotes proliferation and inhibits apoptosis of in vitro cultured buffalo granulosa cells by regulation the expression pattern of associated genes,which indicated that it took an important role in the follicular development of buffalo,and laid a theoretical foundation to clarify the livestock follicular genesis.

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