Department of OBGYN, School of Medicine, University of Pittsburgh, PA 15213, USA.
Mol Hum Reprod. 2013 Jun;19(6):388-94. doi: 10.1093/molehr/gat008. Epub 2013 Jan 31.
Oligozoospermia (low sperm count) is a common semen deficiency. However, to date, few genetic defects have been identified to cause this condition. Moreover, even fewer molecular genetic diagnostic tests are available for patients with oligozoospermia in the andrology clinic. Based on animal and gene expression studies of oligozoospermia, several molecular pathways may be disrupted in post-meiotic spermatozoa. One of the disrupted pathways is protein ubiquitination and cell apoptosis. A critical protein involved in this pathway is the ubiquitin-conjugating enzyme 2B, UBE2B. Absence of Ube2b in male mice causes spermatogenic meiotic disruption with increased apoptosis, leading to infertility. To examine the association between messenger RNA defects in UBE2B and severe oligozoospermia (0.1-10 × 10(6) cells/ml), sequencing of sperm cDNA in 326 oligozoospermic patients and 421 normozoospermic men was performed. mRNA alterations in UBE2B were identified in sperm in 4.6% (15 out of 326) of the oligozoospermic patients, but not found in control men, suggesting strong association between mRNA defects and oligozoospermia (χ(2) = 19, P = 0.0001). Identified UBE2B alterations include nine splicing, four missense and two nonsense alterations. The follow-up screen of corresponding DNA regions did not reveal causative DNA mutations, suggesting a post-transcriptional nature of identified defects. None of these variants were reported in the dbSNP database, although other splicing abnormalities with low level of expression were present in 11 out of 421 (2.6%) controls. Our findings suggest that two distinct molecular mechanisms, mRNA editing and splicing processing, are disrupted in oligozoospermia. We speculate that the contribution of post-transcriptional mRNA defects to oligozoospermia could be greater than previously anticipated.
少精子症(精子数量低)是一种常见的精液缺乏症。然而,迄今为止,只有少数遗传缺陷被确定可导致这种情况。此外,在男科诊所,患有少精子症的患者可使用的分子遗传诊断测试更少。基于对少精子症的动物和基因表达研究,几个分子途径可能在减数分裂后精子中受到干扰。其中一个受干扰的途径是蛋白质泛素化和细胞凋亡。在这个途径中涉及的一个关键蛋白是泛素结合酶 2B,UBE2B。雄性小鼠中 Ube2b 的缺失会导致精子发生减数分裂破坏,凋亡增加,导致不育。为了研究 UBE2B 信使 RNA 缺陷与严重少精子症(0.1-10×10(6)细胞/ml)之间的关系,对 326 名少精子症患者和 421 名正常精子症男性的精子 cDNA 进行了测序。在 4.6%(15/326)的少精子症患者的精子中发现了 UBE2B 的 mRNA 改变,但在对照组男性中未发现,这表明 mRNA 缺陷与少精子症之间存在很强的关联(χ(2) = 19,P = 0.0001)。鉴定的 UBE2B 改变包括 9 个剪接、4 个错义和 2 个无义改变。相应 DNA 区域的后续筛查未发现致病 DNA 突变,表明鉴定的缺陷具有转录后性质。这些变体都未在 dbSNP 数据库中报道,尽管在 11 名(2.6%)对照中存在其他低表达的剪接异常。我们的研究结果表明,在少精子症中,有两种不同的分子机制,mRNA 编辑和剪接加工受到干扰。我们推测,转录后 mRNA 缺陷对少精子症的贡献可能比以前预期的更大。