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

doi:10.11843/j.issn.0366-6964.2024.09.020

摘要/Abstract

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。

关键词: 绵羊, FecB突变, 卵泡, BMP/SMAD通路, SMAD4

Abstract:

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 mutant sheep follicles, the activity of the BMP/SMAD pathway was significantly enhanced.

Key words: sheep, FecB mutation, follicle, BMP/SMAD pathway, SMAD4

中图分类号:

S826.3

龚一鸣, 贾一轩, 李佳骏, 王翔宇, 贺小云, 储明星, 狄冉. 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[J]. 畜牧兽医学报, 2024, 55(9): 3957-3967.

Yiming GONG, Yixuan JIA, Jiajun LI, Xiangyu WANG, Xiaoyun HE, Mingxing CHU, Ran DI. BMP/SMAD Pathway Activity and Protein Expression Profiles in Ovarian Follicles with Different Diameters in Diverse FecB Genotyped Ewes[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 3957-3967.

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分组 Category	基因型 Genotype	卵巢个数 Amount of ovary	卵泡直径(平均值±标准差) Diameter of ovarian follicle (Average ±Standard deviation)	每只羊采样卵泡数量 Amount of ovarian follicle collected from each sheep
大卵泡组	FecB野生型	12	6.00±0.74^a	直径>5 mm的1个
Large follicle group	FecB突变型	12	4.40±0.42^b	直径>4 mm的2个
小卵泡组	FecB野生型	12	2.42±0.19	直径 < 3 mm的5个以上
Small follicle group	FecB突变型	12	2.52±0.18	直径 < 3 mm的5个以上

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