酶发光法：用于体外实时监测天然神经毒素及原代皮质神经元释放L-谷氨酸的工具。
Enzyme-luminescence method: Tool for real-time monitoring of natural neurotoxins in vitro and l-glutamate release from primary cortical neurons.
作者信息
Zakir Hossain S M
机构信息
Department of Chemical Engineering, University of Bahrain, P.O. Box 32038, Bahrain.
出版信息
Biotechnol Rep (Amst). 2016 Jan 18;9:57-65. doi: 10.1016/j.btre.2015.12.002. eCollection 2016 Mar.
Novel enzyme-luminescence method is used for the rapid and sensitive in vitro detection of natural neurotoxins (e.g., shellfish and mushroom toxins) using model brain cells. Paralytic shellfish poisons gonyautoxins (e.g., GTX2,3 and GTX1,4) were detected at 1 nM level by their inhibition of glutamate release from C6 glioma cells upon drug stimulation (IC: GTX2,3 = 30 nM and GTX1,4 = 8 nM). Activation of glutamate release from C6 cells by ibotenic acid (a mushroom toxin) was also evaluated (EC = 10 nM). The method was tested for real-time detection of glutamate release from primary rat cortical neurons. Dose-dependent effects of KCl (0-200 mM) and NMDA on glutamate release from primary cortical neurons were studied. The effects of different culture conditions on K-depolarization-induced glutamate release were also investigated. The method may be applicable to screening of drugs and toxins, and finding glutamatergic neurons in brain slices without in situ staining.
新型酶发光法用于利用模型脑细胞快速、灵敏地体外检测天然神经毒素（如贝类和蘑菇毒素）。通过在药物刺激后抑制C6胶质瘤细胞释放谷氨酸，可检测到1 nM水平的麻痹性贝类毒素膝沟藻毒素（如GTX2,3和GTX1,4）（IC：GTX2,3 = 30 nM，GTX1,4 = 8 nM）。还评估了鹅膏蕈氨酸（一种蘑菇毒素）对C6细胞谷氨酸释放的激活作用（EC = 10 nM）。该方法用于实时检测原代大鼠皮层神经元的谷氨酸释放。研究了KCl（0 - 200 mM）和NMDA对原代皮层神经元谷氨酸释放的剂量依赖性影响。还研究了不同培养条件对K⁺去极化诱导的谷氨酸释放的影响。该方法可能适用于药物和毒素的筛选，以及在不进行原位染色的情况下在脑片中寻找谷氨酸能神经元。
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