中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (49): 9169-9173.doi: 10.3969/j.issn.1673-8225.2011.49.011

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

神经干细胞向神经元前体细胞分化过程中NMDA受体介导的电流变化

王湘臻1，2，李  梅3，胡莹莹1，于滋超4，王庆明2，徐铁军1   

  1. 徐州医学院，1解剖学教研室，3神经生物学教研室，江苏省徐州市221002；2江苏省沭阳县人民医院烧伤科，江苏省沭阳县  223600；4徐州医学院附属医院骨科, 江苏省徐州市 221002
  • 收稿日期:2011-05-04 修回日期:2011-06-23 出版日期:2011-12-03 发布日期:2011-12-03
  • 通讯作者: 徐铁军，博士，教授，徐州医学院解剖学教研室，江苏省徐州市 221002 wxz8311019@163.com
  • 作者简介:王湘臻★，男，1983年生，山东省即墨市人，汉族，2011年徐州医学院毕业，硕士，主要从事神经系统的损伤与修复的研究。 wangxiangzhen0613@163.com
  • 基金资助:

    江苏省自然科学基金(BK2009087)。

Development of current mediated by NMDA receptors during differentiation of neural stem cells into neuronal precursor cells

Wang Xiang-zhen1,2, Li Mei3, Hu Ying-ying1, Yu Zi-chao4, Wang Qing-ming2, Xu Tie-jun1   

  1. 1Department of Anatomy, Xuzhou Medical College, Xuzhou  221002, Jiangsu Province, China; 2Department of Burn, Shuyang People’s Hospital, Shuyang  223600, Jiangsu Province, China; 3Department of Neurobiology, Xuzhou Medical College, Xuzhou  221002, Jiangsu Province, China; 4Department of Orthopaedics, Affiliated Hospital of Xuzhou Medical College, Xuzhou  221002, Jiangsu Province, China
  • Received:2011-05-04 Revised:2011-06-23 Online:2011-12-03 Published:2011-12-03
  • Contact: Xu Tie-jun, Doctor, Professor, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China wxz8311019@163.com
  • About author:Wang Xiang-zhen★, Master, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China wangxiangzhen0613@163.com
  • Supported by:

    the Natural Science Foundation of Jiangsu Province, No. BK2009087*

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摘要：

背景：NMDA受体是脑内主要的兴奋性氨基酸谷氨酸的特异性受体，参与中枢神经系统许多重要的生理和病理过程。
目的：观察体外培养的SD大鼠海马区的神经干细胞及神经前体细胞在发育过程中NMDA受体介导的全细胞电流的变化情况。
方法：应用细胞免疫化学方法观察神经干细胞的nestin以及分化1，3，7 d 神经前体细胞的βⅢ-tubulin表达情况。应用全细胞膜片钳技术在-60 mV钳制电压下记录神经干细胞和神经前体细胞电流。将神经干细胞和神经前体细胞和分为对照组和实验组，对照组加入无Mg2+的细胞外液；实验组加入10，20，50，100 μmol/L NMDA。
结果与结论：神经干细胞的NMDA受体未检测到介导电流产生；分化1，2 d的神经元前体细胞未检测到电流；分化3，7 d的神经元前体细胞能够检测到电流，随着NMDA剂量的增加电流也在增大；分化7 d的神经元前体细胞在相同剂量的NMDA诱导产生的电流较分化3 d的产生的电流大。提示：①实验中未检测到神经干细胞的NMDA介导电流产生。②神经前体细胞第3天时检测到NMDA介导电流。 ③随着神经前体细胞分化的进展，NMDA介导电流在增大。

关键词: 神经元前体细胞, 神经干细胞, NMDA受体, 膜片钳技术, 电生理

Abstract:

BACKGROUND: PNMDA receptors are major specific excitatory amino acid glutamate receptors in the brain. Studies have shown that NMDA receptors are involved in many important physiological and pathological processes of the central nervous system.
OBJECTIVE: To determine the NMDA receptor-mediated whole-cell current changes during the development of neural stem cells (NSCs) and neural precursor cells (NPCs) in SD rat hippocampus in vitro.
METHODS: NPCs were induced for 1, 3, 7 and 9 days. Nestin and βⅢ-tubulin positive cells were assessed by immunocytochemistry. Currents were recorded at -60 mV using patch clamp techniques. The NSCs and NPCs were divided into control and experimental groups. For the control group, cells were perfused with Mg2+-free extracellular solution. For the experimental group, cells were respectively perfused with 10, 20, 50 and 100 μmol/L NMDA.
RESULTS AND COUCLUSION: There were no NMDA receptor-induced currents recorded in NSCs. No NMDA receptor-induced currents were detected in NPCs after 1-2 days of differentiation. Current records were obtained after 3 and 7 days of differentiation, and the currents were strengthened with the increasing dose of NMDA. The currents obtained after 7 days of differentiation were larger than those obtained after 3 days of differentiation at the same dose of NMDA. It was indicated that: ①NMDA-mediated currents in NSCs were not detected during experiments. ②NMDA-mediated currents in NPCs were detected after 3 days of differentiation. ③With the differentiation progress of NPCs, NMDA-mediated currents increased.

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