Major seminal plasma proteome of rabbits and associations with sperm quality
Major seminal plasma proteome of rabbits and associations with sperm quality
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DOI：
10.1016/j.theriogenology.2019.01.013
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发表时间：
2019-04-01
期刊：
影响因子：
2.8
通讯作者：
Moura, A. A.
中科院分区：
文献类型：
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作者：
Bezerra, M. J. B.;Arruda-Alencar, J. M.;Moura, A. A.
本研究描述了兔精浆的主要蛋白质组，以及精浆蛋白质与精液标准之间的潜在联系。采集18只新西兰成年兔精液，采用双向凝胶电泳法和串联质谱仪分析精浆蛋白。观察精子活动率、活力、浓度、形态和膜精子存活率。兔射精量为364+/-7000万精子/ml，活动细胞占81+/-6.1%，活力3.8+/-0.2，形态正常的精子占66.7+/-2.5%。根据精子存活率和顶体完整性分析，65.8+/-2.5%的活精子顶体完整，大多数精子顶体和功能膜均完整。根据PDQuest软件(美国Bio Rad)的测定，兔精浆的二维凝胶平均有232+/-69.5个斑点。质谱学可以鉴定出137种不同的蛋白质。兔精浆中含量最丰富的蛋白质是血红蛋白亚基Zeta样蛋白、膜联蛋白、Lipocalin、FAM115蛋白和白蛋白。与这五个蛋白质相关的斑点的强度代表了在主凝胶中检测到的所有斑点强度的71.5%。以精子性状为因变量，精浆蛋白为自变量，建立多元回归模型。精子活力与β-神经生长因子和富含半胱氨酸的分泌蛋白-1呈正相关，与半乳糖凝集素-1呈负相关。兔精子膜完好率与精浆蛋白FAM115复合体和原肌球蛋白有关。因此，兔精液中形态正常的精子数量与癌胚抗原相关细胞黏附分子6呈正相关，并受精浆异柠檬酸脱氢酶的下调。根据另一个回归模型，部分解释了活精子顶体完整百分率的变化是由精液中白细胞弹性酶抑制剂和肽基脯氨酰顺反异构酶A的数量所解释的。本研究对兔精浆中的137种蛋白质进行了鉴定。精液中的主要蛋白质主要涉及防止脂质过氧化自由基和氧化应激造成的损伤、膜功能、脂质向精子膜的运输和温度调节。此外，找到精浆蛋白作为精液参数的指示器将会改进辅助生殖技术。(C)2019 Elsevier Inc.保留所有权利。
The present study was conducted to describe the major seminal plasma proteome of rabbits and potential associations between seminal proteins and semen criteria. Semen samples were collected from 18 New Zealand adult rabbits, and seminal plasma proteins were analyzed by 2-D SDS-PAGE and tandem mass spectrometry. Sperm motility, vigor, concentration, morphology and membrane sperm viability were evaluated. Rabbits ejaculated 364 +/- 70 million sperm/ml, with 81 +/- 6.1% motile cells, 3.8 +/- 0.2 vigor and 66.7 +/- 2.5% sperm with normal morphology. Based on the viability and acrosome integrity assay, there were 65.8 +/- 2.5% live sperm with intact acrosome and most spermatozoa had both intact acrosome and functional membrane. On average, 2-D gels of rabbit seminal plasma had 232 +/- 69.5 spots, as determined by PDQuest software (Bio Rad, USA). Mass spectrometry allowed the identification of 137 different proteins. The most abundant proteins in rabbit seminal plasma were hemoglobin subunit zeta like, annexins, lipocalin, FAM115 protein and albumin. The intensity of the spots associated with these five proteins represented 71.5% of the intensity of all spots detected in the master gel. Multiple regression models were estimated using sperm traits as dependent variables and seminal plasma proteins as independent ones. Also, sperm motility had positive association with beta-nerve growth factor and cysteine-rich secretory protein 1-like and a negative one with galectin-1. The percentage of rabbit sperm with intact membrane was related to seminal plasma protein FAM115 complex and tropomyosin. Then, the population of morphologically normal sperm in rabbit semen was positively linked to carcinoembryonic antigen-related cell adhesion molecule 6-like and down regulated by seminal plasma isocitrate dehydrogenase. Based on another regression model, the variation in the percentage of live sperm with intact acrosome was partially explained by the amount of leukocyte elastase inhibitor and the peptidylprolyl cis-trans isomerase A in the rabbit seminal fluid. The current study reports the identification of 137 proteins of rabbit seminal plasma. Major proteins of seminal secretion relate primarily to prevention of damages caused by lipid peroxide radicals and oxidative stress, membrane functionality, transport of lipids to the sperm membrane and temperature regulation. Moreover, finding seminal plasma proteins as indicators of semen parameters will improve assisted reproductive technologies. (C) 2019 Elsevier Inc. All rights reserved.