解剖学报 ›› 2022, Vol. 53 ›› Issue (5): 545-550.doi: 10.16098/j.issn.0529-1356.2022.05.001

• 神经生物学 •    下一篇

共济失调毛细血管扩张突变基因在维持小鼠海马神经干细胞静息状态中的作用

董传明* 何文华   

  1. 南通大学医学院人体解剖学系,江苏省神经再生重点实验室,神经再生协同创新中心,江苏 南通 226001
  • 收稿日期:2022-04-08 修回日期:2022-05-30 出版日期:2022-10-06 发布日期:2022-10-06
  • 通讯作者: 董传明 E-mail:yiyimarket@163.com
  • 基金资助:
    国家自然科学基金面上项目

Role of ataxia-telangiectasia mutated gene in maintaining quiescence of neural stem cell from subgranular zone of mouse hippocampus

DONG  Chuan-ming*  HE  Wen-hua#br#   

  1. Department of Human Anatomy,Medical College of Nantong University, Jiangsu Key Laboratory of Nerve Regeneration, Collaborative Innovation Center of Nerve Regeneration, Jiangsu Nantong 226001,China
  • Received:2022-04-08 Revised:2022-05-30 Online:2022-10-06 Published:2022-10-06
  • Contact: DONG Chuan-ming E-mail:yiyimarket@163.com

摘要:

目的  探讨共济失调毛细血管扩张突变基因(ATM)在维持小鼠海马齿状回颗粒下层(SGZ)神经干细胞(NSCs)静息状态中的作用。   方法  构建ATM基因敲除小鼠,1月龄和4月龄ATM基因敲除小鼠各12只,体外分离、培养、鉴定ATM基因敲除小鼠SGZ 区NSCs,Nestin/ki67染色比较1月龄ATM+/+和ATM-/-小鼠SGZ区神经干细胞的增殖水平;体内用BrdU染色比较1月龄和4月龄小鼠SGZ区细胞的增殖水平,胶质纤维酸性蛋白(GFAP)/性别决定基因高迁移率组蛋白2(SOX2)检测4月龄两组小鼠SGZ区静息神经干细胞比例的差异。   结果  1月龄时,体外Nestin/ki67染色和体内BrdU染色均显示ATM-/-小鼠SGZ区神经干细胞表现出异常高的增殖能力(P<0.05)。4月龄时,体内BrdU染色显示,ATM-/-小鼠SGZ区神经干细胞的增殖能力明显降低(P<0.01),GFAP/SOX2染色显示ATM-/-小鼠SGZ区静息神经干细胞的比例明显降低(P<0.01)。   结论  ATM的缺失使NSCs不能维持静息状态,导致NSCs短暂性大量扩增,继而出现NSCs库的耗竭。提示ATM对维持神经干细胞静息起到重要作用。

关键词: 共济失调毛细血管扩张突变基因, 神经干细胞, 静息, 免疫荧光, 小鼠

Abstract:

Objective  To investigate the role of ataxia-telangiectasia mutated gene (ATM) in maintaining quiescence of neural stem cells (NSCs) from subgranular zone (SGZ) of hippocampal dentate gyrus in mice.    Methods  We constructed 1-month-old and 4-month-old mice ATM knockout mice, with 12 mice in each group. The NSCs in SGZ of ATM knockout mice were isolated, cultured and identified in vitro. The proliferation ability of NSCs in SGZ of 1-month-old ATM+/+ and ATM-/- mice were compared by Nestin/ki67 staining; BrdU staining was used to compare the proliferation ability of cells between the two groups in 1-month-old and 4-month-old mice in SGZ, and glial fibrillary acidic protein (GFAP)/sex determining region Y-box 2 (SOX2) staining was used to detect the difference in the proportion of quiescent NSCs between the two groups in SGZ of 4-month-old mice.    Results   Abnormal increased proliferation of NSCs in 1 month-old ATM-/-mice detected by Nestin/ki67 staining in vitro and BrdU staining in vivo(P<0.05). At the age of 4 months, BrdU staining in vivo showed that the proliferation of NSCs in the SGZ of ATM-/- mice decreased significantly(P<0.01), and GFAP/SOX2 staining showed that the proportion of quiescent NSCs in SGZ of ATM-/- mice decreased significantly(P<0.01).  Conclusion  ATM deficiency reduces NSCs quiescence, thus leading to increased activation, proliferation and eventual exhaustion of NSCs, suggesting that ATM can maintain the neural stem cell quiescence. 

Key words: Ataxia-telangiectasia mutated gene, Neural stem cell, Quiescence, Immunofluorescence, Mouse

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