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Proteomic Markers of Functional Sperm Population in Bovines: Comparison of Low- and High-Density Spermatozoa Following Cryopreservation
Journal of Proteome Research ( IF 4.4 ) Pub Date : 2017-11-28 00:00:00 , DOI: 10.1021/acs.jproteome.7b00493
Olivier D’Amours , Gilles Frenette , Sylvie Bourassa , Ézéchiel Calvo , Patrick Blondin 1 , Robert Sullivan
Affiliation  

Mammalian semen contains a heterogeneous population of sperm cells. This heterogeneity results from variability in the complex processes of cell differentiation in the testis, biochemical modifications undergone by spermatozoa during transit along the male reproductive tract, interactions with secretions from accessory sex glands at ejaculation, and, in the context of reproductive technologies, in the ability of ejaculated spermatozoa to resist damage associated with freeze–thaw procedures. When submitted to density gradient centrifugation, ejaculated spermatozoa distribute themselves into two distinct populations: a low-density population characterized by low motility parameters, and a high-density population with high motility characteristics. To understand the origin of ejaculated spermatozoa heterogeneity, cryopreserved semen samples from bulls used by the artificial insemination (A.I.) industry were submitted to Percoll gradient centrifugation. Proteins from low and high density spermatozoa were then extracted with sodium deoxycholate and submitted to proteomic analysis using iTRAQ (isobaric tag for relative and absolute quantitation) methodologies. Quantification of selected sperm proteins was confirmed by multiple reaction monitoring (MRM). Overall, 31 different proteins were more abundant in low-density spermatozoa, while 80 different proteins were more abundant in the high-density subpopulation. Proteins enriched in high-density spermatozoa were markers of sperm functionality such as the glycolytic process, binding to the egg zona pellucida, and motility. Low-density spermatozoa were not solely characterized by loss of proteins and their associated functions. Chaperonin-containing TCP1s and chaperones are hallmarks of the low-density subpopulation. iTRAQ analysis revealed that other proteins such as binder of sperm proteins, histone, GPX5, ELSPBP1, and clusterin are overexpressed in low-density spermatozoa suggesting that these proteins represent defects occurring at different steps during the sperm journey. These differences contribute to the sperm cell heterogeneity present in mammalian semen.

中文翻译:

牛功能性精子种群的蛋白质组学标记:低温保存后低密度和高密度精子的比较

哺乳动物的精液含有异质的精子细胞群。这种异质性是由于睾丸中细胞分化的复杂过程的可变性,精子沿着男性生殖道转运过程中精子经历的生化修饰,射精时与性腺分泌物的相互作用以及生殖技术方面的原因所致。射精的精子具有抵抗冻融过程相关损害的能力。当进行密度梯度离心时,射精的精子将自己分为两个不同的种群:以低运动参数为特征的低密度种群,以及具有高运动特征的高密度种群。要了解射精的异质性的起源,人工授精(AI)行业使用的公牛冷冻保存的精液样本已提交Percoll梯度离心。然后用脱氧胆酸钠提取低密度和高密度精子中的蛋白质,并使用iTRAQ(等压标记用于相对和绝对定量)对蛋白质组学进行分析。通过多反应监测(MRM)确认了所选精子蛋白的定量。总体而言,低密度精子中31种不同蛋白含量更高,而高密度亚群中80种不同蛋白含量更高。富含高密度精子的蛋白质是精子功能的标志物,例如糖酵解过程,与卵透明带的结合以及运动性。低密度精子不仅仅以蛋白质及其相关功能的丧失为特征。包含伴侣蛋白的TCP1和伴侣蛋白是低密度亚人群的标志。iTRAQ分析显示,其他蛋白,例如精子蛋白,组蛋白,GPX5,ELSPBP1和簇蛋白的结合蛋白在低密度精子中过表达,表明这些蛋白代表了在精子旅程中不同步骤发生的缺陷。这些差异有助于哺乳动物精液中存在的精子细胞异质性。在低密度精子细胞中过表达α-淀粉和簇蛋白的现象表明这些蛋白质代表了在精子旅程中不同步骤发生的缺陷。这些差异有助于哺乳动物精液中存在的精子细胞异质性。在低密度精子细胞中过表达α-淀粉和簇蛋白的现象表明这些蛋白质代表了在精子旅程中不同步骤发生的缺陷。这些差异有助于哺乳动物精液中存在的精子细胞异质性。
更新日期:2017-11-29
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