低温处理对猪精子泛素化水平及体外受精的影响

发布时间：2019-02-26 08:24

【摘要】：本研究的主要目的是探索低温处理对猪精子蛋白泛素化水平及体外受精的影响。本试验主要采用CASA、SDS-PAGE、Western blot等试验方法进行分析,得出如下结果：1.常温、冷休克、低温处理后附睾精子活率、顶体完整性还是质膜完整性均高于精子试验组。2.常温、冷休克、低温处理后附睾精子的平均曲线运动速度(VCL)、平均直线运动速度(VSL)、平均路径运动速度(VAP)均高于精子试验组。3.常温、冷休克、低温处理后鲜精和附睾精子的蛋白进行SDS-PAGE,实验结果显示,冷休克和低温后的猪精子蛋白含量或种类发生了变化,而鲜精与附睾精子样本基本没有差异。4.冷休克处理的猪精子样本进行Western blot分析。实验结果表明：得知约在30kDa、46kDa和62kDa处存在泛素分子：对46kDa处进行灰度分析,各处理组之间条带的灰度值比都差异明显；62kDa处只在鲜精和附睾精子中发现泛素分子；而冷休克处理的泛素化程度相对其它两处理组降低很多。5.低温处理的猪精子样本进行Western blot分析。实验结果表明：得知约在46kDa和62kDa处存在泛素分子,对46kDa处的泛素化程度进行灰度分析,精子-80℃处理组与其它三个处理组之间的灰度比值都小很多；附睾精子-196℃处理组与其它三个处理组之间的灰度比值都大很多。6.冷休克处理的猪精子与成熟卵母细胞进行体外受精,受精2d后观察卵裂情况,并统计卵裂率。实验结果表明：附睾鲜精卵裂率高于鲜精；冷休克附睾精子卵裂率高于冷休克精子。7.低温处理的猪精子与成熟卵母细胞进行体外受精,受精2d后观察卵裂情况,并统计卵裂率。实验结果表明：附睾精子-80℃卵裂率高于精子-80℃；附睾精子-196℃的卵裂率高于精子-196℃。
[Abstract]:The aim of this study was to explore the effects of low temperature treatment on the level of ubiquitin and in vitro fertilization of porcine sperm. The results of this experiment are as follows: 1. The results are as follows: 1. CASA,SDS-PAGE,Western blot and other test methods are used for analysis. The sperm motility, acrosome integrity or plasma membrane integrity of epididymal spermatozoa after normal temperature, cold shock and hypothermic treatment were higher than those of sperm treatment group. 2. The mean curvilinear velocity (VCL),) of epididymal sperm after normal temperature, cold shock and hypothermic treatment was higher than that of sperm treated with low temperature (VSL), mean linear velocity (VSL), mean path velocity (VAP). The results of SDS-PAGE, test of fresh sperm and epididymal spermatozoa after cold shock and cold shock showed that the protein content or species of pig spermatozoa changed after cold shock and hypothermic treatment. There was no difference between fresh sperm and epididymal sperm samples. 4. Cold shock-treated pig sperm samples were analyzed by Western blot. The results showed that there were ubiquitin molecules at about 30kDas, 46kDa and 62kDa. The gray-scale ratio of the bands at 46kDa was significantly different among the treatment groups, and only ubiquitin molecules were found in fresh sperm and epididymal sperm in 62kDa, while ubiquitin molecules were only found in fresh sperm and epididymal spermatozoa in 62kDa. The degree of ubiquitin of cold shock treatment was much lower than that of the other two groups. 5. 5%. Low temperature-treated pig sperm samples were analyzed by Western blot. The results showed that there were ubiquitin molecules in 46kDa and 62kDa, and the degree of ubiquitin at 46kDa was analyzed by gray scale analysis. The gray scale ratio between sperm-80 鈩，

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