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对公猪精浆蛋白进行蛋白质分析,这些蛋白在精子低温保存过程中具有保护作用。

Protein analysis of boar seminal plasma proteins with protective effect during low-temperature storage of spermatozoa.

作者信息

Daskalova Denica, Kukov Alexander, Kirilova Irina, Ivanova-Kicheva Maria

机构信息

Department of Reproductive Biotechnology and Cryobiology of Gametes, Institute of Biology and Immunology of Reproduction, Bulgarian Academy of Sciences , Sofia , Bulgaria.

出版信息

Biotechnol Biotechnol Equip. 2014 Jul 4;28(4):716-720. doi: 10.1080/13102818.2014.926679. Epub 2014 Oct 28.

Abstract

The present study aimed to investigate the effect of different seminal plasma proteins (SPPs) on boar spermatozoa functional characteristics. We investigated the putative protective effect of SSPs on sperm cells motility and velocity, as well as on the integrity of the plasma membrane (PM) during low-temperature storage at 4 °C. SPPs fractions were obtained and purified by gel permeation chromatography (GPC). Nine fractions of SPPs were obtained and further characterized by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Sperm computer analysis (SCA) after incubation of spermatozoa with separated proteins revealed that fraction 6 consisting of low molecular weight (MW) proteins could preserve spermatozoa motility and velocity better when compared to those with higher MW. Two-dimensional (2D) elecrtophoretic analysis showed that fraction 6 contained proteins with the following MW and isoelectric point (pI): 16 kDa and pI 7.35, 18 kDa and pI 5.20, 19 kDa and pI 7.35, 26 kDa and pI 4.50, 26 kDa and pI 4.30, 29 kDa and pI 5.85.

摘要

本研究旨在探讨不同精浆蛋白(SPPs)对公猪精子功能特性的影响。我们研究了SPPs在4℃低温保存期间对精子细胞活力和速度以及质膜(PM)完整性的假定保护作用。通过凝胶渗透色谱法(GPC)获得并纯化了SPPs组分。获得了九个SPPs组分,并通过12%十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)进一步表征。精子与分离出的蛋白质孵育后的精子计算机分析(SCA)显示,与高分子量的组分相比,由低分子量(MW)蛋白质组成的组分6能更好地保持精子活力和速度。二维(2D)电泳分析表明,组分6包含以下分子量和等电点(pI)的蛋白质:16 kDa和pI 7.35、18 kDa和pI 5.20、19 kDa和pI 7.35、26 kDa和pI 4.50、26 kDa和pI 4.30、29 kDa和pI 5.85。

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