荣昌猪BMP15基因通过TGF-β RⅡ激活SMAD4信号分子机制研究

doi:10.11843/j.issn.0366-6964.2020.04.011

Abstract

Abstract: In order to clarify BMP15 gene expression characteristics in Rongchang pigs at different developmental stages in relation to their sexual maturity, as well as the relationship between the expression of the BMP15 gene and SMADs signaling pathways genes, ovarian tissues from 1 month, 3 months, and 5 months old Rongchang pigs were collected to analyze different BMP15 gene expression and cellular localization characteristics in ovarian tissues of the Rongchang pigs at different developmental stages using qRT-PCR and paraffin sectioning methods. BMP15 and SMADs signaling pathways related genes SMAD2, SMAD3, SMAD4, SMAD7, TGF-β1, TGF-β2, TGF-β3, TGF-β RⅠ and TGF-β RⅡ expression characteristics were also revealed using qRT-PCR and Western blot analysis by adding recombinant human BMP15 protein and TGF-β receptor inhibitor (LY215799 and LY2109761) to 5-month-old ovarian biopsy. The results showed that from the age of 1 month to 5 months, the mRNA expression of BMP15 gene in ovarian tissues were up-regulated (P<0.05)with the growth and development of Rongchang pigs. Immunofluorescence and paraffin sectioning experiments showed that BMP15 protein fluorescent signals existd in granulosa cells surrounding oocyte in the 5-month-old ovarian tissues. Within the ovarian tissues, from 3-5 months old, BMP15, SMAD4, TGF-β1 and TGF-β RⅡ genes mRNA expressions were raised and SMAD2, TGF-β2 and TGF-β RⅠ expressions reduced (P<0.05). By adding the re combinant human BMP15 protein, TGF-β RⅠ/ Ⅱ and TGF-β RⅠ receptor inhibitor in the age of 5 months old ovarian biopsy, the results showed that TGF-β RⅠ/ Ⅱ receptor inhibitor (LY2109761) significantly inhibited the expression of TGF-β RⅡ proteins and TGF-β RⅠ receptor inhibitor (LY2157299) could not inhibit the expression of TGF-β RⅡ proteins. In conclusion, BMP15 gene expression before sexual maturity in Rongchang pig ovarian tissues were raised and SMAD4, TGF-β1 and TGF-β RⅡ genes expression were also up-regulated. The study illuminated that BMP15 gene mediated SMAD4 signaling molecules and regulated the downstream genes expression through TGF-β R Ⅱ, which provided theoretical basis for the further study of Rongchang pig BMP15 gene roles in the follicles development.

Key words: Rongchang pig, BMP15 gene, SMAD, oocyte, TGF-β

CLC Number:

S828.8

YI Huashan, MA Xianping, ZHAO Yao, ZHU Piao, LIU Xiao, DENG Boyuan, WU Xinwei, ZHU Dan, CUI Yan. Study on the Rongchang Pig BMP15 Gene through TGF-β RⅡ Activation SMAD4 Signal Molecular Mechanism[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 754-762.

References 25

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Study on the Rongchang Pig BMP15 Gene through TGF-β RⅡ Activation SMAD4 Signal Molecular Mechanism

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