Volume 11, Issue 6 (November 2009) 11, 683–693; 10.1038/aja.2009.59
Proteome analysis of round-headed and normal spermatozoa by 2-D fluorescence difference gel electrophoresis and mass spectrometry
Ting-Ting Liao1,*, Zhen Xiang1,*, Wen-Bing Zhu1 and Li-Qing Fan1
Institute of Human Reproduction and Stem Cell Engineering, Central South University, Changsha 410078, China
* Ting-Ting Liao and Zhen Xiang contributed equally to this work.
Correspondence: Prof. Li-Qing Fan, fanliqingszzx@sina.com.cn
Received 18 April 2009; Revised 7 July 2009; Accepted 3 September 2009; Published online 12 October 2009.
Abstract |
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. Keywords: differential protein, globozoospermia, mass spectrometry (MS), two-dimensional difference gel electrophoresis (2-D DIGE)
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