摘要
目的研究无镁诱导对惊厥后发育中皮层神经元表达γ-氨基丁酸A受体（GABAAR）α1亚单位的影响。方法以原代培养的大鼠大脑皮层神经元无镁诱导的反复惊厥样放电为模型，根据对培养6d皮层神经元的不同处理分为正常细胞外液组和无镁细胞外液组。神经元在上述2种液体中孵育3h，然后恢复正常DMEM培养液继续培养，1、7及12d后应用流式细胞技术、免疫细胞化学技术及实时荧光定量PCR测定2组大鼠皮层神经元GABAARα1亚单位表达的变化。结果应用流式细胞技术发现无镁损伤后12d时无镁组GABAARα1阳性细胞率较对照组明显下降，差异有统计学意义（P〈0．05）；应用免疫细胞化学方法发现无镁损伤后7d、12d时实验组GABAARα1蛋白表达较对照组明显下降，差异有统计学意R（P〈0．05）；应用实时荧光定量PCR发现无镁损伤后1d时，无镁组GABAARα1mRNA表达较对照组明显下降，差异有统计学意义（P〈0．05）。结论早期无镁细胞外液处理可以诱导发育中大鼠皮层神经元GABAARα1表达的改变。
Objective To study the expression of gamma-aminobutyric acid type A receptor （GABAAR） α1 subunit following convulsion induced by magnesium-free medium in developing cortical neurons. Methods Rat embryo cortical neurons cultured for 6 days were cultured for 3 h in normal or magnesium-free extracellular fluid, followed by further culture in DMEM. At 1, 7 and 12 days after magnesium-free treatment, RT-PCR, immunocytochemical staining, and flow cytometry were performed to detect the expression of GABAAR α1 subunit. Results Compared with that in the control group, GABAAR α1-positive cells in the magnesium-free group decreased significantly at 12 days （P〈0.05）. Immunocytochemical staining showed a significant decrease of accumulated optical density （AOD） of GABAAR ctl subunit in the magnesium-free group at 7 and 12 days as compared with that in the control group （P〈0.05）; the mRNA expression ofGABAAR α1 subunit in the magnesium-free group decreased significantly at 1 day compared to that in the control group （P〈0.05）. Conclusion The expression of GABAAR α1 subunit is altered following convulsion induced by magnesium-free medium in developing cortical neurons.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2009年第11期1090-1093,共4页
Chinese Journal of Neuromedicine
基金
国家自然科学基金青年科学基金（30400483）
关键词
惊厥
Γ-氨基丁酸A受体
α1亚单位
神经元
发育
Convulsion
Type A gamma-aminobutyric acid receptor
α1 subunit
Neurons
Development
