中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (14): 1297-1306.doi: 10.3969/j.issn.1673-5374.2013.14.006

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

分化使HT22神经元对兴奋性毒性更敏感

  

  • 收稿日期:2012-12-13 修回日期:2013-02-23 出版日期:2013-05-15 发布日期:2013-05-15

Differentiation renders susceptibility to excitotoxicity in HT22 neurons

Minchao He1, 2, Jun Liu1, 2, Shaowu Cheng2, Yigang Xing1, William Z Suo2, 3, 4   

  1. 1 Department of Neurology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China
    2 Laboratory for Alzheimer’s Disease & Aging Research, Veterans Affairs Medical Center, Kansas, MO 64128, USA
    3 Department of Neurology, University of Kansas Medical Center, Kansas, KS 66170, USA
    4 Molecular & Integrative Physiology, University of Kansas Medical Center, Kansas, KS 66170, USA
  • Received:2012-12-13 Revised:2013-02-23 Online:2013-05-15 Published:2013-05-15
  • Contact: William Z Suo, M.D., Associate professor, Laboratory for Alzheimer’s Disease & Aging Research, Veterans Affairs Medical Center, Kansas, MO 64128, USA; Departments of Neurology, University of Kansas Medical Center, Kansas, KS 66170, USA; Molecular & Integrative Physiology, University of Kansas Medical Center, Kansas, KS 66170, USA, William.Suo@va.gov.
  • About author:Minchao He☆, M.D.
  • Supported by:

    研究由美国医学研究和发展服务部，退伍军人事务部和中西部地区生物医学研究基金会，和阿尔茨海默氏症协会（NPSPAD-11-202149）进行资助。

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摘要：

HT22是一个永生化小鼠海马神经元细胞系，它不像成熟的海马神经元能在体内表达胆碱能和谷氨酸受体。在与记忆相关的研究中，这种特性在某种程度上阻碍了其作为一个成熟的海马神经元模型的使用。文章报告在体内对HT22细胞进行适当的分化诱导，使其变成和保持类似于那些成熟海马神经元的属性，如变得对谷氨酸更易兴奋。实验发现谷氨酸诱导毒性的敏感性在未分化和已分化的HT22细胞有显著差异，已分化HT22细胞的EC50降低约两个数量级。此外，N-甲基-D-天冬氨酸受体拮抗剂MK-801和美金刚均能减少谷氨酸对已分化HT22细胞的毒性，而不能减少未分化HT22细胞的毒性。显然，已分化HT22细胞可表达N-甲基-D-天门冬氨酸受体，未分化的HT22细胞不能。这些结果表明，分化对永生化细胞系呈现有丝分裂后的神经元特性是重要的，而且与未分化的HT22细胞相比，已分化HT22神经元是海马神经元一个更好的模型。

关键词: 神经再生, 脑损伤, HT22, 分化, N-甲基-D-天门冬氨酸受体, 谷氨酸, 兴奋性神经毒性, 有丝分裂, 海马, 神经元, MK-801, 美金刚, 基金资助文章

Abstract:

HT22 is an immortalized mouse hippocampal neuronal cell line that does not express cholinergic and glutamate receptors like mature hippocampal neurons in vivo. This in part prevents its use as a model for mature hippocampal neurons in memory-related studies. We now report that HT22 cells were appropriately induced to differentiate and possess properties similar to those of mature hippocampal neurons in vivo, such as becoming more glutamate-receptive and excitatory. Results showed that sensitivity of HT22 cells to glutamate-induced toxicity changed dramatically when comparing undifferentiated with differentiated cells, with the half-effective concentration for differentiated cells reducing approximately two orders of magnitude. Moreover, glutamate-induced toxicity in differentiated cells, but not undifferentiated cells, was inhibited by the N-methyl-D- aspartate receptor antagonists MK-801 and memantine. Evidently, differentiated HT22 cells expressed N-methyl-D-aspartate receptors, while undifferentiated cells did not. Our experimental findings indicated that differentiation is important for immortalized cell lines to render post-mitotic neuronal properties, and that differentiated HT22 neurons represent a better model of hippocampal neurons than undifferentiated cells.

Key words: neural regeneration, brain injury, HT22, differentiation, N-methyl-D-aspartate receptor, glutamate, excitatory neurotoxicity, mitosis, hippocampus, neurons, MK-801, memantine, grants-supported paper, neuroregeneration