Uysal Fatma, Akkoyunlu Gokhan, Ozturk Saffet
Department of Histology and Embryology, Akdeniz University School of Medicine, Campus, 07070, Antalya, Turkey; and Department of Histology and Embryology, Ankara University School of Medicine, 06100, Ankara, Turkey.
Department of Histology and Embryology, Akdeniz University School of Medicine, Campus, 07070, Antalya, Turkey.
Reprod Fertil Dev. 2019 Jul;31(8):1386-1394. doi: 10.1071/RD18246.
DNA methylation plays key roles in epigenetic regulation during mammalian spermatogenesis. DNA methyltransferases (DNMTs) function in de novo and maintenance methylation processes by adding a methyl group to the fifth carbon atom of the cytosine residues within cytosine-phosphate-guanine (CpG) and non-CpG dinucleotide sites. Azoospermia is one of the main causes of male infertility, and is classified as obstructive (OA) or non-obstructive (NOA) azoospermia based on histopathological characteristics. The molecular background of NOA is still largely unknown. DNA methylation performed by DNMTs is implicated in the transcriptional regulation of spermatogenesis-related genes. The aim of the present study was to evaluate the cellular localisation and expression levels of the DNMT1, DNMT3A and DNMT3B proteins, as well as global DNA methylation profiles in testicular biopsy samples obtained from men with various types of NOA, including hypospermatogenesis (hyposperm), round spermatid (RS) arrest, spermatocyte (SC) arrest and Sertoli cell-only (SCO) syndrome. In the testicular biopsy samples, DNMT1 expression and global DNA methylation levels decreased gradually from the hyposperm to SCO groups (P P P <0.05). Although both DNMT1 and DNMT3A were localised in the cytoplasm and nucleus of the spermatogenic cells, staining for DNMT3B was more intensive in the nucleus of spermatogenic cells. In conclusion, the findings suggest that significant changes in DNMT expression and global DNA methylation levels in spermatogenic cells may contribute to development of male infertility in the NOA groups. Further studies are needed to determine the molecular biological effects of the altered DNMT expression and DNA methylation levels on development of male infertility.
DNA甲基化在哺乳动物精子发生过程的表观遗传调控中发挥着关键作用。DNA甲基转移酶(DNMTs)通过在胞嘧啶-磷酸-鸟嘌呤(CpG)和非CpG二核苷酸位点的胞嘧啶残基的第五个碳原子上添加甲基,在从头甲基化和维持甲基化过程中发挥作用。无精子症是男性不育的主要原因之一,根据组织病理学特征可分为梗阻性(OA)或非梗阻性(NOA)无精子症。NOA的分子背景在很大程度上仍然未知。由DNMTs执行的DNA甲基化与精子发生相关基因的转录调控有关。本研究的目的是评估DNMT1、DNMT3A和DNMT3B蛋白的细胞定位和表达水平,以及从患有各种类型NOA的男性获得的睾丸活检样本中的整体DNA甲基化谱,包括精子发生低下(hyposperm)、圆形精子细胞(RS)阻滞、精母细胞(SC)阻滞和唯支持细胞(SCO)综合征。在睾丸活检样本中,从精子发生低下组到SCO组,DNMT1表达和整体DNA甲基化水平逐渐降低(P P P<0.05)。虽然DNMT1和DNMT3A都定位于生精细胞的细胞质和细胞核中,但DNMT3B在生精细胞核中的染色更强。总之,这些发现表明生精细胞中DNMT表达和整体DNA甲基化水平的显著变化可能有助于NOA组男性不育的发生。需要进一步研究以确定DNMT表达和DNA甲基化水平改变对男性不育发生的分子生物学影响。