不同FecB基因型和不同直径绵羊卵泡中BMP/SMAD通路活性及蛋白表达差异  

BMP/SMAD Pathway Activity and Protein Expression Profiles in Ovarian Follicles with Different Diameters in Diverse FecB Genotyped Ewes

作　　者：龚一鸣 贾一轩 李佳骏 王翔宇[1] 贺小云[1] 储明星[1] 狄冉[1] GONG Yiming;JIA Yixuan;LI Jiajun;WANG Xiangyu;HE Xiaoyun;CHU Mingxing;DI Ran(State Key Laboratory of Animal Biotech Breeding,Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,畜禽生物育种全国重点实验室,北京100193

出　　处：《畜牧兽医学报》2024年第9期3957-3967,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31861143012);财政部和农业农村部:国家现代农业产业技术体系(CARS-38);中国农业科学院科技创新工程(CAAS-ZDRW202106,ASTIP-IAS13)。

摘　　要：旨在探究不同FecB基因型对绵羊卵泡中BMP/SMAD通路活性和蛋白表达的影响;揭示成熟大卵泡和小卵泡之间BMP/SMAD通路活性和蛋白表达的差异。本研究采用TaqMan分型方法筛选出不同FecB基因型的母羊,同期发情后取卵泡期成熟卵泡和黄体期卵巢表面小卵泡,利用免疫印迹法(Western blot)测定BMP/SMAD通路相关蛋白表达水平和通路活性。结果表明,对于小卵泡组,FecB突变型卵泡中骨形态发生蛋白1B型受体(bone morphogenetic protein receptor type 1B,BMPR1B)表达量显著高于野生型卵泡(P<0.05),但SMAD家族成员4(SMAD family member 4,SMAD4)表达量和SMAD1/5/9的磷酸化水平显著低于野生型卵泡(P<0.05);对于成熟大卵泡组,FecB突变型卵泡中FKBP脯氨酰异构酶1A(FKBP prolyl isomerase 1A,FKBP1A)和SMAD4表达量显著低于野生型卵泡(P<0.05),Ⅰ型受体(BMPR1B)和Ⅱ型受体(BMPR2)的蛋白表达量及SMAD1/5/9的磷酸化水平在两种基因型之间未显示出显著差异。另一方面,对比FecB突变型小卵泡和成熟大卵泡发现:成熟大卵泡中BMPR1B和SMAD4蛋白表达量和SMAD1/5/9磷酸化程度显著高于小卵泡(P<0.05)。上述结果表明,由于SMAD4表达量的下降,FecB突变型大、小卵泡中结合到基因组靶区域的SMAD4-SMAD1/5/9蛋白复合物均相对较少,将导致通路活性降低,而且由于小卵泡中较低的SMAD1/5/9磷酸化水平,其通路活性更低。另外,绵羊突变型卵泡生长发育成熟后BMP/SMAD通路活性显著增强。This study aimed to investigate the effects of different FecB genotypes on the BMP/SMAD signaling pathway activity and protein expression in sheep follicles,and explored the differences in BMP/SMAD signaling pathway activity and protein expression between mature large follicles and small follicles.Firstly,TaqMan method was used to screen ewes with different genotypes of FecB.Secondly,after estrus synchronization,mature follicles in follicular phase and small follicles in luteal phase were sampled,and the pathway activity and the protein expression of BMP/SMAD pathway were determined by Western blot.The result showed that,for the small follicle group,the expression of bone morphogenetic protein receptor type 1B(BMPR1B)in FecB mutant follicles was significantly higher than that in wild-type follicles(P<0.05),but the expression of SMAD family member 4(SMAD4)and the phosphorylation levels of SMAD1/5/9 were significantly lower than those in wild-type follicles(P<0.05).For the mature large follicle group,the expression of FKBP prolyl isomerase 1A(FKBP1A)and SMAD4 in FecB mutant follicles was significantly lower than that in wild-type follicles(P<0.05),and there was no significant difference in the protein expression of type I receptor(BMPR1B)and type II receptor(BMPR2)and the phosphorylation levels of SMAD1/5/9 between the two genotypes.On the other hand,when comparing FecB mutant small follicles with mature large follicles,the protein expression levels of BMPR1B and SMAD4 and the phosphorylation level of SMAD1/5/9 in mature large follicles were significantly higher than those in small follicles(P<0.05).The results indicated that due to the decrease in the expression of SMAD4,the FecB mutant large and small follicles have relatively fewer SMAD4-SMAD1/5/9 protein complexes bound to the genomic target regions,which means a reduction in pathway activity.Moreover,due to the lower phosphorylation level of SMAD1/5/9 in small follicles,its pathway activity was lower.Additionally,after the development and maturation of the mut

关 键 词：绵羊 FecB突变 卵泡 BMP/SMAD通路 SMAD4 

分 类 号：S826.3[农业科学—畜牧学]