Hu J, el-Fakahany E E
Division of Neuroscience Research in Psychiatry, University of Minnesota Medical School, Minneapolis 55455.
Life Sci. 1994;55(13):1009-16. doi: 10.1016/0024-3205(94)00635-0.
It is important to apply an appropriate test for determining cell viability, in order to properly evaluate the role of the amyloid beta protein in neuronal degeneration in Alzheimer's disease. In the current paper, we present evidence that the putative neurotoxic fragment 25-35 of amyloid beta causes loss of trypan blue exclusion in differentiated mouse neuroblastoma N1E-115 cells which suggests a potential neurotoxic effect. Surprisingly, no parallel changes in apparent cell viability were observed when fluorescein diacetate staining or release of lactate dehydrogenase were measured. Positive staining with trypan blue was also induced by incubating cell membranes prepared from N1E-115 cells or rat hippocampus with amyloid beta 25-35. Our results indicate that amyloid beta might induce trypan blue adsorption on the cell membrane. Therefore, caution should be taken when trypan blue exclusion is used in studies of the potential neurotoxicity of amyloid beta peptides.
为了正确评估淀粉样β蛋白在阿尔茨海默病神经元变性中的作用，应用适当的测试来确定细胞活力很重要。在当前论文中，我们提供证据表明，淀粉样β假定的神经毒性片段25 - 35导致分化的小鼠神经母细胞瘤N1E - 115细胞中锥虫蓝排斥丧失，这表明存在潜在的神经毒性作用。令人惊讶的是，当测量荧光素二乙酸酯染色或乳酸脱氢酶释放时，未观察到明显细胞活力的平行变化。用淀粉样β 25 - 35孵育由N1E - 115细胞或大鼠海马体制备的细胞膜也可诱导锥虫蓝阳性染色。我们的结果表明，淀粉样β可能诱导锥虫蓝吸附在细胞膜上。因此，在使用锥虫蓝排斥研究淀粉样β肽的潜在神经毒性时应谨慎。
