| 摘要: |
| 【目的】探索热应激蛋白90（HSP90）表达量与牛精液冷冻-解冻后品质的关系。【方法】采用假阴道法采集身体健康、年龄2～4岁的9头荷斯坦奶牛精液（编号1～9），测定新鲜精液指标（活力、形态、活率和密度），评定其品质，合格的精液样品经过冷冻 解冻后检测其精子活率、质膜完整率和顶体完整率，并根据冻后品质将其分为HFTs(High freezing resistance teams)和LFTs（Low freezing resistance teams) 2组，对精子HSP90蛋白进行SDS-聚丙烯酰胺凝胶（SDS-PAGE）电泳和Western blot检测，分析其表达量与精子抗冻性的关系。【结果】不同牛个体精液冻后品质相差较大。SDS-PAGE电泳结果显示，HFTs组的分子质量约为90 ku的蛋白表达量显著高于LFTs组（P＜0.05），经Western blot分析，该蛋白为HSP90。HSP90蛋白表达量与冷冻-解冻后的牛精子品质呈明显正相关，其与精子活率、顶体完整率、质膜完整率的相关系数分别为0.364，0.447和0.402。【结论】HSP90蛋白表达量可以作为判断牛精子抗冻性的指标之一。 |
| 关键词: 牛 精液品质 HSP90表达量 抗冻性 |
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| 基金项目:科技部成果转化项目（2010GB2G000466）；教育部长江学者与创新团队支持计划项目(IRT0940)；陕西省农业攻关项目（2011K01-04）；陕西省“13115”科技创新工程项目（2010ZDGC-01）；陕西省农业科技创新项目(2011NXC01-11)；西北农林科技大学基本科研业务费专项（PY200901） |
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| Correlation study on HSP90 expression and bull sperm freezing resistance |
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| Abstract: |
| 【Objective】The present study was conducted to investigate fertility-associated proteins in bull sperm and the correlation between 90 ku heat-shock protein (HSP90) and semen characteristics.【Method】Semens of 9 Holstein cow were collected by a bovine artificial vagina.The fresh semen characteristics of sperm motility,morphology,viability and concentration were evaluated.After semens were freeze-thawed,sperm progressive motility,membrane integrity and acrosome integrity were assessed.Then the 9 ejaculates were clustered for freezing resistance on the basis of their sperm quality,and sperm protein was subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and Western blot.【Result】Sperm quality assessment proved that individual differences of sperm quality after frozen were large.The SDS-PAGE results revealed that there was a substantial diverse of 90 ku protein in frozen-thawed spermatozoa. Western blot analysis demonstrated that the protein was HSP90.Thus,this study indicated that the expression of HSP90 was associated with semen characteristics.Related coefficient between HSP90 expression and sperm motility,membrane integrity,acrosome integrity respectively was 0.364,0.447 and 0.402.【Conclusion】The expression of HSP90 could be utilized in developing a reliable and simple method to determine the bull sperm freezing resistance. |
| Key words: bull sperm characteristics HSP90 expression freezing resistance |
