Wang Xiangyu, Guo Xiaofei, He Xiaoyun, Di Ran, Zhang Xiaosheng, Zhang Jinlong, Chu Mingxing
State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology, Tianjin Engineering Research Center of Animal Healthy Farming, Institute of Animal Science and Veterinary, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China.
Animals (Basel). 2023 Dec 19;14(1):11. doi: 10.3390/ani14010011.
The fecundity () mutation in the bone morphogenetic protein receptor type 1B (BMPR1B) gene increases ovulation in sheep. However, its effect on follicular maturation is not fully understood. Therefore, we collected granulosa cells (GCs) at a critical stage of follicle maturation from nine wild-type (), nine heterozygous mutant (), and nine homozygous mutant () Small Tail Han sheep. The GCs of three ewes were selected at random from each genotype and consolidated into a single group, yielding a total of nine groups (three groups per genotype) for proteomic analysis. The tandem mass tag technique was utilized to ascertain the specific proteins linked to multiple ovulation in the various genotypes. Using a general linear model, we identified 199 proteins significantly affected by the mutation with the LIMMA package ( < 0.05). The differential abundance of proteins was enriched in pathways related to cholesterol metabolism, carbohydrate metabolism, amino acid biosynthesis, and glutathione metabolism. These pathways are involved in important processes for GC-regulated 'conservation' of oocyte maturation. Further, the sparse partial least-squares discriminant analysis and the Fuzzy-C-mean clustering method were combined to estimate weights and cluster differential abundance proteins according to ovulation to screen important ovulation-related proteins. Among them, ZP2 and ZP3 were found to be enriched in the cellular component catalog term "egg coat", as well as some apolipoproteins, such as APOA1, APOA2, and APOA4, enriched in several Gene Ontology terms related to cholesterol metabolism and lipoprotein transport. A higher abundance of these essential proteins for oocyte maturation was observed in BB and WB genotypes compared with WW ewes. These proteins had a high weight in the model for discriminating sheep with different genotypes. These findings provide new insight that the mutant in GCs improves nutrient metabolism, leading to better oocyte maturation by altering the abundance of important proteins (ZP2, ZP3, and APOA1) in favor of increased ovulation or better oocyte quality.
骨形态发生蛋白受体1B（BMPR1B）基因中的繁殖力（）突变可增加绵羊的排卵量。然而，其对卵泡成熟的影响尚未完全明确。因此，我们从9只野生型（）、9只杂合突变型（）和9只纯合突变型（）小尾寒羊的卵泡成熟关键阶段采集了颗粒细胞（GCs）。从每种基因型中随机选取3只母羊的GCs合并为一组，共得到9组（每种基因型3组）用于蛋白质组学分析。采用串联质量标签技术确定与不同基因型多次排卵相关的特定蛋白质。使用通用线性模型，我们通过LIMMA软件包鉴定出199种受突变显著影响的蛋白质（<0.05）。蛋白质的差异丰度富集在与胆固醇代谢、碳水化合物代谢、氨基酸生物合成和谷胱甘肽代谢相关的途径中。这些途径参与了颗粒细胞调节卵母细胞成熟“保守”的重要过程。此外，结合稀疏偏最小二乘判别分析和模糊C均值聚类方法，根据排卵情况估计权重并对差异丰度蛋白质进行聚类，以筛选重要的排卵相关蛋白质。其中，ZP2和ZP3在细胞成分目录术语“卵壳”中富集，一些载脂蛋白，如APOA1、APOA2和APOA4，在与胆固醇代谢和脂蛋白转运相关的几个基因本体术语中富集。与WW母羊相比，在BB和WB基因型中观察到这些卵母细胞成熟必需蛋白质的丰度更高。这些蛋白质在区分不同基因型绵羊的模型中具有较高权重。这些发现提供了新的见解，即颗粒细胞中的突变改善了营养代谢，通过改变重要蛋白质（ZP2、ZP3和APOA1）的丰度，有利于增加排卵或提高卵母细胞质量，从而实现更好的卵母细胞成熟。
