Rolseth Veslemøy, Rundén-Pran Elise, Luna Luisa, McMurray Cynthia, Bjørås Magnar, Ottersen Ole Petter
Centre for Molecular Biology and Neuroscience, Institute of Medical Microbiology, University of Oslo, Rikshospitalet HF, Oslo, Norway.
DNA Repair (Amst). 2008 Sep 1;7(9):1578-88. doi: 10.1016/j.dnarep.2008.06.007. Epub 2008 Jul 22.
High metabolic activity and low levels of antioxidant enzymes make neurons particularly prone to damage by reactive oxygen species. Thus, repair of oxidative DNA damage is essential for normal brain function. Base excision repair is the major pathway for repair of oxidative DNA damage, and is initiated by DNA glycosylases recognizing and removing the damaged base. In mammalian cells at least five different DNA glycosylases with overlapping substrate specificity, NEIL1, NEIL2, NEIL3, OGG1 and NTH1, remove oxidative DNA base lesions. Here we report mRNA expression and distribution of these five DNA glycosylases in human and rodent brains using in situ hybridization and Northern blotting supported by glycosylase activity assays. NEIL1, NEIL2, OGG1 and NTH1 showed widespread expression at all ages. In situ hybridization studies in mouse brain showed that expression of mNeil1 increased with age. In newborn mouse brain, mNeil3 revealed a discrete expression pattern in brain regions known to harbour stem cell populations, i.e., the subventricular zone, the rostral migratory stream, and the hilar region of the hippocampal formation. Expression of mNeil3 decreased with age, and in old mice brains could be detected only in layer V of neocortex. MNth1 was constitutively expressed during lifespan. In Northern blots, mOgg1 expression showed a transient decrease followed by an increase after 8 weeks of age. Assays for faPy DNA glycosylase activity revealed increased activity level with age in all brain regions analyzed. The widespread but differential expression of the DNA glycosylases recognizing oxidative base lesions suggests distinct and age dependent roles of these enzymes in genome maintenance in brain. The distribution of mNeil3 is particularly intriguing and points to a specific role of this enzyme in stem cell differentiation.
高代谢活性和低水平的抗氧化酶使神经元特别容易受到活性氧的损伤。因此,氧化DNA损伤的修复对于正常脑功能至关重要。碱基切除修复是氧化DNA损伤修复的主要途径,由识别并去除受损碱基的DNA糖基化酶启动。在哺乳动物细胞中,至少有五种具有重叠底物特异性的不同DNA糖基化酶,即NEIL1、NEIL2、NEIL3、OGG1和NTH1,可去除氧化DNA碱基损伤。在此,我们通过原位杂交和Northern印迹法,并辅以糖基化酶活性测定,报告这五种DNA糖基化酶在人和啮齿动物大脑中的mRNA表达及分布情况。NEIL1、NEIL2、OGG1和NTH1在所有年龄段均广泛表达。对小鼠脑进行的原位杂交研究表明,mNeil1的表达随年龄增长而增加。在新生小鼠脑中,mNeil3在已知含有干细胞群体的脑区,即脑室下区、嘴侧迁移流和海马结构的齿状回门区,呈现离散的表达模式。mNeil3的表达随年龄下降,在老年小鼠脑中仅在新皮质第V层可检测到。MNth1在整个生命周期中持续表达。在Northern印迹中,mOgg1的表达在8周龄时短暂下降,随后上升。对faPy DNA糖基化酶活性的测定显示,在所有分析的脑区中,活性水平均随年龄增加。识别氧化碱基损伤的DNA糖基化酶的广泛但有差异的表达表明,这些酶在大脑基因组维持中具有不同的、依赖年龄的作用。mNeil3的分布特别引人关注,表明该酶在干细胞分化中具有特定作用。