小脑颗粒神经元谷氨酸兴奋性毒性模型的优化

发布时间：2018-03-05 19:36

本文选题：兴奋性毒性　切入点：小脑颗粒神经元　出处：《生物化学与生物物理进展》2013年08期 　论文类型：期刊论文

【摘要】：在急性、慢性神经退行性疾病和炎症引发的神经系统疾病的发病机制中,兴奋性毒性可能是造成后期神经元死亡的共同途径.小脑颗粒神经元谷氨酸兴奋性毒性模型是研究上述过程的重要实验手段,该模型的稳定性和可重复性是开展相关研究的重要基础.然而,文献报道的建模方法条件各异,说法不一,很难适从.本工作针对小脑颗粒神经元谷氨酸兴奋性毒性模型建立的关键环节,包括小脑颗粒神经元的培养、兴奋性毒性刺激条件的确定,毒性标志性指标的表征,分别进行了比较和优化,从培养皿的包被、神经元消化、兴奋性刺激的溶液介质选择、神经元刺激的最佳时间及谷氨酸的最佳刺激浓度等方面分别给出了优化条件.通过特征性钙离子曲线、NMDA受体特异性抑制剂MK-801的干预作用以及c-fos基因转录水平的动力学变化等指标,确认了毒性模型的成功建立.本工作不仅对建立小脑颗粒神经元谷氨酸兴奋性毒性模型的实验室具有重要参考意义,而且,其针对不同条件分析比较的结果及优化原则,对其他神经毒性模型的建立也具有普遍参考意义.
[Abstract]:In the pathogenesis of acute, chronic neurodegenerative and inflammatory neurological diseases, Excitatory toxicity may be the common pathway of neuronal death in late stage. The model of glutamate excitotoxicity in cerebellar granular neurons is an important experimental means to study the above process. The stability and repeatability of the model is an important basis for the relevant research. However, the modeling methods reported in the literature are different in terms and conditions. This work aims at the key links of the establishment of glutamate excitatory toxicity model of cerebellar granule neurons, including the culture of cerebellar granule neurons, the determination of excitatory toxicity stimulation conditions, the characterization of the iconic toxic indexes, The experiments were compared and optimized from the culture dish coating, neuronal digestion, excitatory stimulation of the solution medium selection, The optimal time of neuronal stimulation and the optimal concentration of glutamic acid were given. The intervention effect of NMDA receptor specific inhibitor MK-801 and the transcriptional level of c-fos gene were analyzed by characteristic calcium curve. Mechanical changes and other indicators, The successful establishment of the toxicity model was confirmed. This work not only has important reference significance for the laboratory of the model of glutamate excitotoxicity in cerebellar granule neurons, but also analyses the results and optimization principles according to different conditions. It is also of general reference significance for the establishment of other neurotoxic models.
【作者单位】：石河子大学医学院;中国科学院生物物理研究所;
【基金】：国家自然科学基金委重点项目(31030023) 石河子大学高层次人才科研启动项目(RCZX201113)~~
【分类号】：R-332

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