解剖学报 ›› 2013, Vol. 44 ›› Issue (6 ): 735-739.doi: 10.3969/j.issn.0529-1356.2013.06.002

• 神经生物学 • 上一篇    下一篇

Lhx8促进海马神经干细胞向胆碱能神经元分化

田美玲 朱培培 金国华* 李浩明 秦建兵 施金洪   

  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,江苏 南通 226001
  • 收稿日期:2012-12-24 修回日期:2013-01-31 出版日期:2013-12-06 发布日期:2013-12-06
  • 通讯作者: 金国华 E-mail:jguohua@ntu.edu.cn
  • 基金资助:

    国家自然科学基金资助项目;省自然科学基金资助项目;自然科学基金资助项目

Lhx8 promotes the differentiation of hippocampal neural stem cells into cholinergic neurons in vitro

TIAN Mei-ling ZHU Pei-pei JIN Guo-hua* LI Hao-ming QIN Jian-bing SHI Jin-hong   

  1. Department of Anatomy, Medical School, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Jiangsu Nantong 226001 china
  • Received:2012-12-24 Revised:2013-01-31 Online:2013-12-06 Published:2013-12-06
  • Contact: JIN Guo-hua E-mail:jguohua@ntu.edu.cn

摘要:

目的 构建大鼠Lhx8全长结构基因真核表达载体。方法 PCR法扩增Lhx8全长结构基因,经EcoRI和BamHI双酶切后连接至真核表达载体pEGFP-C3中,重组质粒pEGFP-C3-Lhx8经测序鉴定后,采用脂质体转染的方法转染至体外培养的海马神经干细胞中。 结果 测序鉴定表明,重组质粒pEGFP-C3- Lhx8-EGFP构建成功,该重组质粒转入体外培养的神经干细胞后,质粒转染组中ChAT阳性的细胞数较阴性对照组明显增多(P<0.05)。 结论 重组质粒pEGFP-C3- Lhx8-EGFP构建成功,Lhx8能够促进NSCs向ChAT阳性的细胞分化。

关键词: Lhx8, 神经干细胞, 海马, 基因表达, 聚合酶链反应, 大鼠

Abstract:

Objective To construct eukaryotic expression vector containing full length cDNA of rat Lhx8 gene. Methods  Full length cDNA of rat Lhx8 gene was acquired by PCR method and digested by EcoRI and BamHI. The digested gene fragment was inserted into eukaryotic expression vector pEGFP-C3 and identified by DNA sequencing. The recombinant plasmid was transfected into hippocampal neural stem cells (NSCs) by liposome. Results DNA sequencing demonstrated that the insertion fragment in recombinant plasmid was right, and recombinant plasmid pEGFP-C3-Lhx8 was transfected into NSCs. After 7 days, the number of ChAT positive cells in plasmid transefection group was significantly higher than the negative control group. Conclusion Eukaryotic expression vector pEGFP-C3- Lhx8 is constructed successfully, and Lhx8 promotes the differentiation of NSCs into ChAT positive cellsin vitro.

Key words: Lhx8, Neural stem cell, Hippocampus, Gene expression, PCR, Rat