Hetherington Louise, Schneider Elena K, Scott Caroline, DeKretser David, Muller Charles H, Hondermarck Hubert, Velkov Tony, Baker Mark A
From the ‡Discipline of Biological Sciences, School of Biomedical Sciences and Pharmacy, Hunter Medical Research Institute,University of Newcastle, Callaghan, NSW 2308, Australia.
§Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Vic, 3052, Australia.
Mol Cell Proteomics. 2016 Dec;15(12):3685-3693. doi: 10.1074/mcp.M116.060343. Epub 2016 Oct 21.
Globally, ∼1 in 15 men of reproductive age are infertile, yet the precise mechanisms underlying their gamete failure are unknown. Although a semen analysis is performed to determine fertilizing potential, the diagnostic suitability of this analysis has been questioned in several reports, as many men, classified as infertile according to their semen analysis, subsequently turn out to be fertile. Herein, we have used a quantitative (phospho)-proteomic analysis, using enrichment on titanium dioxide followed by ion-trap mass spectrometry (LC-MS/MS), to compare the semen of infertile versus fertile males. One protein, namely outer dense fiber 1 (ODF1), was dramatically reduced in infertile males. Using specific antibodies, we then screened the gametes of a cohort of suspected infertile men and demonstrated a reduction in the amount of ODF1 compared with fertile controls. Stress treatment of sperm deficient in ODF1 caused the head to decapitate, suggesting why these gametes fail to initiate fertilization. Interestingly, electron micrographs of ODF1-deficient spermatozoa revealed an abnormal connecting piece, indicating several developmental defects with both the implantation plate and the thin laminated fibers. In some cases, the implantation plate appeared to be reduced in size or was overburdened by granular material near the connecting piece. Hence, a strong reduction ODF1 is a marker of idiopathic male infertility and a potential driver of this condition.
在全球范围内,约每15名育龄男性中就有1人不育,然而其配子功能衰竭的确切机制尚不清楚。尽管会进行精液分析以确定受精潜力,但该分析的诊断适用性在几份报告中受到质疑,因为许多根据精液分析被归类为不育的男性后来被证明是可育的。在此,我们使用了定量(磷酸化)蛋白质组分析,通过二氧化钛富集,然后进行离子阱质谱分析(液相色谱-串联质谱法),来比较不育男性与可育男性的精液。一种名为外致密纤维1(ODF1)的蛋白质在不育男性中显著减少。然后,我们使用特异性抗体对一组疑似不育男性的配子进行筛选,结果表明与可育对照组相比,ODF1的含量有所减少。对缺乏ODF1的精子进行应激处理会导致头部脱离,这表明了这些配子无法启动受精的原因。有趣的是,缺乏ODF1的精子的电子显微镜图像显示连接段异常,表明植入板和薄层纤维均存在一些发育缺陷。在某些情况下,植入板似乎尺寸减小,或者在连接段附近被颗粒物质覆盖。因此,ODF1的大量减少是特发性男性不育的一个标志,也是这种情况的一个潜在驱动因素。