进行本研究是为了比较杂交牛和Zebu牛的生精细胞的蛋白质组学特征,以了解杂交公牛亚生育力发生率较高的可能原因。通过细针抽吸从青春期前(6个月)和成年(24个月)杂交和zebu雄性睾丸中收集的生精细胞在体外增殖,蛋白质组学分析是使用鸟枪蛋白质组学方法完成的。在生精细胞中鉴定了蛋白质表达水平的年龄和物种特异性变化。在青春期前的zebu和杂交中鉴定出的差异表达蛋白(DEP)数量为546,而在成年zebu和杂交公牛之间鉴定出579种DEP。在这些中,194个DEPS在这些组中是常见的,40个DEP显示倍数变化≥2。然而,在青春期前以及成虫和杂交公牛中,只有20种蛋白质表现出相似的表达变化趋势(上调或下调)。在这20个部门中,13种蛋白质上调,与杂交公牛相比,zebu的生精细胞中7种蛋白质下调。上调的蛋白质包括RPLP2,PAXIP1,calumenin,prosaposin,GTF2F1,TMP2,泛素结合因子E4A,COL1A2波形蛋白,FAM13A蛋白,外周素,GFPT2和GRP78。与杂交相比,在斑马公牛中下调的7种蛋白质包括APOA1,含G补丁域的蛋白质1,NADP转氨酶线粒体,谷氨酰氨基肽酶,突触素1片段,具有SH3结构域ANK重复和含PH结构域的蛋白1和蛋白转运蛋白sec16B的ArfGAP。推断与精子功能和受精过程相关的蛋白质,比如calumenin,prosaposin,波形蛋白,可以进一步研究GRP78和APOA1,以了解杂交公牛不育的确切原因。
The present study was carried out to compare the proteomic profiles of spermatogenic cells of crossbred and zebu cattle in an effort to understand the possible reasons for a higher incidence of sub-fertility in crossbred bulls. The spermatogenic cells collected from the testes of pre-pubertal (6 mo) and adult (24 mo) crossbred and zebu males through fine needle aspiration were proliferated in vitro, and proteomic profiling was done using a shotgun proteomics approach. The age- and species-specific variations in the expression level of proteins were identified in spermatogenic cells. The number of differentially expressed proteins (DEPs) identified in pre-pubertal zebu and crossbred was 546, while 579 DEPs were identified between adult zebu and crossbred bulls. Out of these, 194 DEPS were common to these groups and 40 DEPs displayed a fold change ≥2. However, only 20 proteins exhibited similar expression variation trends (upregulated or downregulated) among pre-pubertal as well as adult zebu and crossbred bulls. Out of these 20 DEPs, 13 proteins were upregulated, and 7 proteins were downregulated in spermatogenic cells of zebu compared to crossbred bulls. Among the upregulated proteins were RPLP2, PAXIP1, calumenin, prosaposin, GTF2F1, TMP2, ubiquitin conjugation factor E4A, COL1A2, vimentin, protein FAM13A, peripherin, GFPT2, and GRP78. Seven proteins that were downregulated in zebu bulls compared to crossbred included APOA1, G patch domain-containing protein 1, NAD P transhydrogenase mitochondrial, glutamyl aminopeptidase, synaptojanin 1 fragment, Arf GAP with SH3 domain ANK repeat and PH domain-containing protein 1, and protein transport protein sec16B. It was inferred that the proteins associated with sperm function and fertilization processes, such as calumenin, prosaposin, vimentin, GRP78, and APOA1 could be studied further to understand the precise cause of subfertility in crossbred bulls.