Ishihara K, Kanai S, Sago H, Yamakawa K, Akiba S
Department of Pathological Biochemistry, Kyoto Pharmaceutical University, 5 Misasagi-Nakauchi-cho, Yamashina-ku, Kyoto 607-8414, Japan; Laboratory for Neurogenetics, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako City, Saitama 351-0198, Japan.
Department of Pathological Biochemistry, Kyoto Pharmaceutical University, 5 Misasagi-Nakauchi-cho, Yamashina-ku, Kyoto 607-8414, Japan.
Neuroscience. 2014 Dec 5;281:1-15. doi: 10.1016/j.neuroscience.2014.09.039. Epub 2014 Sep 28.
To identify molecular candidates involved in brain disabilities of Ts1Cje, a mouse model of Down syndrome (DS), we performed comparative proteomic analyses. Proteins extracted from the brains of postnatal wild-type (WT) and Ts1Cje mice were analyzed by two-dimensional gel electrophoresis (2-DE). No differences were detected in the proteins expressed in the whole brain between WT and Ts1Cje mice at postnatal day 0 and 3months of age. Five spots with differential expression in the brains of Ts1Cje mice were detected by 2-DE of brain proteins from WT and Ts1Cje embryos at embryonic day 14.5 (E14.5). These differentially expressed proteins in Ts1Cje embryos were identified as calcyclin-binding protein (CACYBP), nucleoside diphosphate kinase-B (NDPK-B), transketolase (TK), pyruvate kinase (PK), and 60S acidic ribosomal protein P0 (RPLP0) by peptide mass fingerprinting. CACYBP and NDPK-B were involved in cell proliferation, whereas TK and PK were associated with energy metabolism. Experiments on cell proliferation, an in vivo bromodeoxyuridine (BrdU)-labeling experiment, and immunohistochemical analysis for phospho-histone H3 (an M-phase marker) demonstrated increased numbers of BrdU-positive and M-phase cells in the ganglionic eminence. Our findings suggest that the dysregulated expression of proteins demonstrated by comparative proteomic analysis could be a factor in increased cell proliferation, which may be associated with abnormalities in DS brain during embryonic life.
为了鉴定唐氏综合征（DS）小鼠模型Ts1Cje脑功能障碍所涉及的分子候选物，我们进行了比较蛋白质组学分析。通过二维凝胶电泳（2-DE）分析从新生野生型（WT）和Ts1Cje小鼠脑中提取的蛋白质。在出生后第0天和3个月大时，WT和Ts1Cje小鼠全脑中表达的蛋白质未检测到差异。在胚胎第14.5天（E14.5），通过对WT和Ts1Cje胚胎脑蛋白进行2-DE，在Ts1Cje小鼠脑中检测到5个差异表达斑点。通过肽质量指纹图谱将Ts1Cje胚胎中这些差异表达的蛋白质鉴定为钙周期蛋白结合蛋白（CACYBP）、核苷二磷酸激酶-B（NDPK-B）、转酮醇酶（TK）、丙酮酸激酶（PK）和60S酸性核糖体蛋白P0（RPLP0）。CACYBP和NDPK-B参与细胞增殖，而TK和PK与能量代谢相关。细胞增殖实验、体内溴脱氧尿苷（BrdU）标记实验以及磷酸化组蛋白H3（M期标记物）的免疫组织化学分析表明，神经节隆起中BrdU阳性细胞和M期细胞数量增加。我们的研究结果表明，比较蛋白质组学分析所显示的蛋白质表达失调可能是细胞增殖增加的一个因素，这可能与胚胎期DS脑的异常有关。
