Choy Yoon-Jung, Hong Sung-Young, Pack Sang-Jin, Woo Ran-Sook, Baik Tai-Kyoung, Song Dae-Yong
Department of Anatomy and Neuroscience, Eulji University School of Medicine, Daejeon, Republic of Korea; Department of Ophthalmology, Eulji University Hospital, Daejeon, Republic of Korea.
Department of Anatomy and Neuroscience, Eulji University School of Medicine, Daejeon, Republic of Korea.
J Chem Neuroanat. 2015 Jul-Sep;66-67:10-8. doi: 10.1016/j.jchemneu.2015.03.003. Epub 2015 Apr 6.
Neuroinflammation is an early event and important contributor to the pathobiology of neurodegenerative diseases. Neuroglia, especially microglia, are a major central nervous system population that can modulate neuroinflammation. To determine potential key molecules in this process, we employed microarray analysis in the substantia nigra (SN) following medial forebrain bundle (MFB) transection and analyzed the temporal expression profiles of candidate genes implicated in neuroglial activation and functional maturation. The DNA microarray analyzed, 8913 probes. Sixty nine genes were up-regulated and 11 genes were down-regulated at least twofold compared to normal control. Of the 80 genes, 23 were related to cell metabolism, 3 related to apoptosis, 27 related to immunity. Among them, 4 genes (Galectin 3, Heat shock protein 27, Lipocalin 2, Tissue inhibitory metalloproteinase 1) seemed to be related to the neuroglial function. The candidate genes were subjected to quantitative real-time PCR, Western blotting, and immunohistochemical approaches. Expression changes similar to the microarray were evident. In a double immunofluorescence assay, Galectin 3 almost completely co-localized with OX6-positive activated microglia, and Heat shock protein 27 mainly co-localized with glial fibrillary acidic protein (GFAP) positive astrocytes. Lipocalin 2, except for a few matches of GFAP positive astrocytes, did not co-localized with any of neuroglial markers. This is the first study to evaluate gene expression changes in the SN following MFB transection, which has been used as a parkinsonian animal model. Several candidate genes with potential roles in neuroglial activation and functional maturation were identified. The molecular significance of the candidate genes in neuroglial activation and neuroinflammation remains unclear.
神经炎症是神经退行性疾病病理生物学过程中的早期事件且是重要促成因素。神经胶质细胞，尤其是小胶质细胞，是可调节神经炎症的主要中枢神经系统细胞群体。为确定此过程中的潜在关键分子，我们在内侧前脑束（MFB）横断后的黑质（SN）中进行了微阵列分析，并分析了与神经胶质细胞激活和功能成熟相关的候选基因的时间表达谱。DNA微阵列分析了8913个探针。与正常对照相比，69个基因上调，11个基因下调至少两倍。在这80个基因中，23个与细胞代谢相关，3个与细胞凋亡相关，27个与免疫相关。其中，4个基因（半乳糖凝集素3、热休克蛋白27、脂质运载蛋白2、组织抑制性金属蛋白酶1）似乎与神经胶质细胞功能相关。对候选基因进行了定量实时PCR、蛋白质印迹和免疫组织化学分析。与微阵列相似的表达变化很明显。在双重免疫荧光分析中，半乳糖凝集素3几乎完全与OX6阳性活化小胶质细胞共定位，热休克蛋白27主要与胶质纤维酸性蛋白（GFAP）阳性星形胶质细胞共定位。脂质运载蛋白2，除了与少数GFAP阳性星形胶质细胞匹配外，未与任何神经胶质细胞标志物共定位。这是第一项评估MFB横断后黑质中基因表达变化的研究，MFB横断已被用作帕金森病动物模型。确定了几个在神经胶质细胞激活和功能成熟中具有潜在作用的候选基因。候选基因在神经胶质细胞激活和神经炎症中的分子意义仍不清楚。
