Effect of cryopreservation on sea bass sperm proteins
Effect of cryopreservation on sea bass sperm proteins
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DOI：
10.1095/biolreprod.104.036202
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发表时间：
2005-05-01
影响因子：
3.6
通讯作者：
Vilella, S
中科院分区：
文献类型：
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作者：
Zilli, L;Schiavone, R;Vilella, S
在本研究中，我们使用二维聚丙烯酰胺凝胶电泳（2-DE）和基质相关激光解吸/电离飞行时间（MALDI-TOF）质谱法来验证海鲈鱼精子的蛋白质表达是否受到低温保存程序的影响。通过目视检查和图像分析软件显示，新鲜和冷冻解冻精液的蛋白质谱不同。我们在新鲜精子中发现了163个斑点；其中13个在冷冻精子二维凝胶中显著减少，8个缺失。其中5个位点用MALDI-TOF进行了分析，但只有3个位点在生物信息学分析的数据库中显示出显著的匹配（Pept-Ident、Mascot和MS-Fit）。其中，斑点5与斑马鱼的一种新蛋白（类似于人和小鼠的SKB1）具有同源性，斑点13与两栖动物G1/ s特异性周期蛋白E2具有同源性，斑点20与Brachidanio rerio的假设蛋白DKFZp566A1524具有同源性。目前的研究表明，使用冷冻保存程序会导致精子蛋白的降解，其中两个可能至少部分地导致观察到的精子运动时间缩短和卵子受精率降低。
In the present study we used two-dimensional polyacryl amide gel electrophoresis (2-DE) and matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to verify whether the protein expression of sea bass sperm was affected by the cryopreservation procedure. The protein profiles differed between fresh and frozen-thawed semen as revealed by visual inspection and by image analysis software. We identified 163 spots in fresh sperm; among these, 13 were significantly decreased and 8 were absent in two-dimensional gel obtained with cryopreserved sperm. Five of these spots were analyzed with MALDI-TOF, but only three showed a significant match in the databases used in bio-informatics analysis (Pept-Ident, Mascot, and MS-Fit). In particular, spot 5 showed homology with a novel protein of zebrafish (similar to SKB1 of human and mouse), spot 13 showed homology with amphibian G1/S-specific cyclin E2, and spot 20 showed homology with the hypothetical protein DKFZp566A1524 of Brachidanio rerio. The present work shows that the use of the cryopreservation procedure causes the degradation of sperm proteins and among these, two could be at least partially responsible for the observed decrease in sperm motility duration and the lower hatching rate of eggs fertilized with cryopreserved sperm.