Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence o...Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence of 100% round-headed spermatozoa on semen analysis, and patients with this condition are absolutely infertile. The objective of this study was to investigate the differences in protein expression between human round- headed and normal spermatozoa. Two-dimensional (2-D) fluorescence difference gel electrophoresis (DIGE) coupled with mass spectrometry (MS) was used in this study. Over 61 protein spots were analysed in each paired normal/round-headed comparison, using DIGE technology along with an internal standard. In total, 35 protein spots identified by tandem mass spectrometry (MS/MS) exhibited significant changes (paired t-test, P 〈 0.05) in the expression level between normal and round-headed spermatozoa. A total of nine proteins were found to be upregulated and 26 proteins were found to be downregulated in round-headed spermatozoa compared with normal spermatozoa. The differentially expressed proteins that we identified may have important roles in a variety of cellular processes and structures, including spermatogenesis, cell skeleton, metabolism and spermatozoa motility.展开更多
目的:探讨双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)的实验方法,分析比较其优缺点,概述其在微生物研究中的应用。方法:以两株不同血清型的副溶血性弧菌为样本,通过2D-DIGE实验,得到两组蛋白质荧光染色...目的:探讨双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)的实验方法,分析比较其优缺点,概述其在微生物研究中的应用。方法:以两株不同血清型的副溶血性弧菌为样本,通过2D-DIGE实验,得到两组蛋白质荧光染色分布图,与银染的效果比较。结果:直观的看到两株细菌的蛋白质差异,且可分析样品丰度变化。结论:该方法有一定的局限性,但却是研究蛋白质组丰度变化的强有力工具。展开更多
基金Acknowledgment We thank Beijing Proteome Research Center, (Beijing, China) for its enthusiastic technological support and for the theory of 2-D DIGE. We also thank(Changsha, China) College of Life Sciences at Hunan Normal University for supporting the MS technology. Finally, we are very grateful to our collaborators for their help, as well as their valuable discussions and suggestions during the course of this work. This work was supported by two grants from the National Natural Science Foundation of China (NO. 30170480 and NO. 30470884).
文摘Globozoospermia is a severe form of teratozoospermia characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. Globozoospermia is diagnosed by the presence of 100% round-headed spermatozoa on semen analysis, and patients with this condition are absolutely infertile. The objective of this study was to investigate the differences in protein expression between human round- headed and normal spermatozoa. Two-dimensional (2-D) fluorescence difference gel electrophoresis (DIGE) coupled with mass spectrometry (MS) was used in this study. Over 61 protein spots were analysed in each paired normal/round-headed comparison, using DIGE technology along with an internal standard. In total, 35 protein spots identified by tandem mass spectrometry (MS/MS) exhibited significant changes (paired t-test, P 〈 0.05) in the expression level between normal and round-headed spermatozoa. A total of nine proteins were found to be upregulated and 26 proteins were found to be downregulated in round-headed spermatozoa compared with normal spermatozoa. The differentially expressed proteins that we identified may have important roles in a variety of cellular processes and structures, including spermatogenesis, cell skeleton, metabolism and spermatozoa motility.
文摘目的:探讨双向差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)的实验方法,分析比较其优缺点,概述其在微生物研究中的应用。方法:以两株不同血清型的副溶血性弧菌为样本,通过2D-DIGE实验,得到两组蛋白质荧光染色分布图,与银染的效果比较。结果:直观的看到两株细菌的蛋白质差异,且可分析样品丰度变化。结论:该方法有一定的局限性,但却是研究蛋白质组丰度变化的强有力工具。