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 (28): 2656-2665.doi: 10.3969/j.issn.1673-5374.2013.28.008

• 9.Original reseatch :nerve repair，regeneration following neural injury • 上一篇    下一篇

神经干细胞向神经元分化过程中表达的2种外向钾电流增加

  

  • 收稿日期:2013-02-07 修回日期:2013-06-08 出版日期:2013-10-05 发布日期:2013-10-05
  • 基金资助:

    国家自然科学基金项目（31000514），新乡医学院高学历人才科研启动项目（No.2007502002）

Two outward potassium current types are expressed during the neural differentiation of neural stem cells

Ruiying Bai1, Guowei Gao2, Ying Xing1, 3, Hong Xue4   

  1. 1 Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China
    2 Department of Radiotherapy, Center Hospital of Xinxiang, Xinxiang 453003, Henan Province, China
    3 Stem Cell Research Center, Zhengzhou University, Zhengzhou 450052, Henan Province, China
    4 Basic Medical Sciences of Henan University of Traditional Chinese Medicine, Zhengzhou 450008, Henan Province, China
  • Received:2013-02-07 Revised:2013-06-08 Online:2013-10-05 Published:2013-10-05
  • Contact: Ying Xing, Professor, Doctoral supervisor, Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China; Stem Cell Research Center, Zhengzhou University, Zhengzhou 450052, Henan Province, China, xingy@zzu.edu.cn.
  • About author:Ruiying Bai, Master, Lecturer.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 31000514, and the Scientific Research Project for Talent with High Education of Xinxiang Medical University, No. 2007502002.

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摘要：

K+通道的电生理特性可作为衡量神经干细胞功能性分化的一项基本指标。实验以此建立新生乳鼠海马神经干细胞的培养体系，采用神经营养生长因子进行诱导，观察其向神经细胞分化过程中K+通道的电生理特性。免疫荧光化学结果显示，神经干细胞在体外能快速增殖形成克隆球并表达高水平的巢蛋白，而分化细胞则表达神经元特异性烯醇化酶。流式细胞仪分析显示， S+G2/M期细胞比例较高，神经干细胞增殖活跃。然而，随细胞分化时间的延长，S+G2/M期细胞比例明显降低。膜片钳记录显示，在细胞发育阶段分化的神经元上K+ 电流有明显的改变，此K+ 电流包括一种快速失活的钾电流成分和一种缓慢失活的钾电流成分，分别能被钾离子通道阻断剂--4-氨基吡啶和四乙铵所阻断。说明新生乳鼠海马神经干细胞在体外一定条件下能被诱导分化为功能性神经元，这些分化的神经元能表达2种类型的外向K+ 电流，从而具有类似体内成熟神经元的基本活性。

关键词: 神经再生, 干细胞, 神经干细胞, 海马, 增殖, 分化, 神经元, 膜片钳, 电生理特性, 外向K+ 电流, 失活, 巢蛋白, 神经元特异性烯醇化酶, 基金资助文章

Abstract:

The electrophysiological properties of potassium ion channels are regarded as a basic index for determining the functional differentiation of neural stem cells. In this study, neural stem cells from the hippocampus of newborn rats were induced to differentiate with neurotrophic growth factor, and the electrophysiological properties of the voltage-gated potassium ion channels were observed. Immunofluorescence staining showed that the rapidly proliferating neural stem cells formed spheres in vitro that expressed high levels of nestin. The differentiated neurons were shown to express neuron-specific enolase. Flow cytometric analysis revealed that the neural stem cells were actively dividing and the percentage of cells in the S + G2/M phase was high. However, the ratio of cells in the S + G2/M phase decreased obviously as differentiation proceeded. Whole-cell patch-clamp re-cordings revealed apparent changes in potassium ion currents as the neurons differentiated. The potassium ion currents consisted of one transient outward potassium ion current and one delayed rectifier potassium ion current, which were blocked by 4-aminopyridine and tetraethylammonium, respectively. The experimental findings indicate that neural stem cells from newborn rat hippo-campus could be cultured and induced to differentiate into functional neurons under defined condi-tions in vitro. The differentiated neurons expressed two types of outward potassium ion currents similar to those of mature neurons in vivo.

Key words: neural regeneration, neural stem cells, hippocampus, proliferation, differentiation, neurons, patch-clamp, electrophysiological properties, transient outward potassium ion current, delayed rec-tifier potassium ion current, inactivation, nestin, neuron-specific enolase, grants-supported paper, neuroregeneration