Identification of biomarker candidates for fertility in spermatozoa of crossbred bulls through comparative proteomics
Identification of biomarker candidates for fertility in spermatozoa of crossbred bulls through comparative proteomics
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DOI：
10.1016/j.theriogenology.2018.06.021
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发表时间：
2018-10-01
期刊：
影响因子：
2.8
通讯作者：
Yadav, Savita
中科院分区：
文献类型：
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作者：
Aslam, M. K. Muhammad;Sharma, Vinay K.;Yadav, Savita
近年来，人们一直在寻找精子中某些蛋白质的表达与生育力之间的关联，以确定男性生育力标记。由于杂交公牛生育力低下的发生率较高，因此本研究对高生育力和低生育力的杂交公牛进行了研究，通过蛋白质组学方法鉴定精子中的生育力标记。使用 2D-DIGE 和 MALDI-TOF-MS 技术将高繁殖力公牛的精子蛋白质组与低繁殖力公牛的精子蛋白质组进行比较，并通过免疫印迹验证结果。 MDH2、ENO1、RIBCI、CAPN7、ATP5D、LacA like Protein-2 like、NCAPD3、DECRI、GCNT2、GDI2、TOP 和 USP12 在高生育力精子中过表达，而 DST like isoform 1、TMEM43 和 BSPI 在低生育力精子中过表达（P < 0.05）。两组之间的差异表达范围为 1.57 (GDI2) 至 5.1 (BSP1) 倍。根据GO注释，它们大多数参与细胞和代谢过程，具有催化和结合活性，并且定位于细胞和细胞器中。其中根据差异表达程度和鉴定可靠性选择ENO1和BSPI进行进一步验证。免疫印迹研究表明ENO1表达与公牛生育力呈正相关（P < 0.05），而BSPI表达与公牛育性呈负相关（P < 0.01）。低生育力公牛冷冻解冻精子中获能精子的比例高于高生育力公牛（P < 0.05）。总的来说，该研究确定了公牛精子中的一些潜在分子，它们可能作为生育力的一组生物标志物。 (C) 2018 Elsevier Inc. 保留所有权利。
Associations between expression of some proteins in spermatozoa and fertility have been sought in recent years to identify the male fertility markers. Since the incidence of sub-fertility is high in crossbred bulls, the present investigation was carried out on high- and low-fertile crossbred bulls to identify fertility markers in spermatozoa through proteomics approach. Sperm proteome of high-fertile bulls were compared with low-fertile bulls using 2D-DIGE and MALDI-TOF-MS techniques and the results were validated with immuno-blotting. The proteins MDH2, ENO1, RIBCI, CAPN7, ATP5D, LacA like protein-2 like, NCAPD3, DECRI, GCNT2, GDI2, TOP and USP12 were over expressed in high-fertile spermatozoa, whereas DST like isoform 1, TMEM43 and BSPI were over expressed in low-fertile spermatozoa (P < 0.05). The differential expression ranged from 1.57 (GDI2) to 5.1 (BSP1) fold between the two groups. Based on the GO annotation, majority of them were involved in cellular and metabolic processes, with catalytic and binding activities, and localized in cell and organelles. Among these proteins, ENO1 and BSPI were selected based on the degree of differential expression and reliability in identification, for further validation. Immuno-blotting studies indicated that ENO1 expression was positively correlated (P < 0.05) while the expression of BSPI was negatively (P < 0.01) correlated with bull fertility. The proportion of capacitated spermatozoa in frozen thawed spermatozoa of low-fertile bulls was higher (P < 0.05) as compared to high-fertile bulls. Collectively, the study identified some potential molecules in spermatozoa of bulls, which may act as a panel of biomarkers for fertility. (C) 2018 Elsevier Inc. All rights reserved.