A novel method for the rapid determination of beta-amyloid toxicity on acute hippocampal slices using MTT and LDH assays
A novel method for the rapid determination of beta-amyloid toxicity on acute hippocampal slices using MTT and LDH assays
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DOI：
10.1016/j.brainresbull.2012.02.005
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发表时间：
2012-04-10
影响因子：
3.8
通讯作者：
Datki, Zsolt
中科院分区：
文献类型：
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作者：
Mozes, Emese;Hunya, Akos;Datki, Zsolt
精确测量β-淀粉样蛋白（A β 1 - 42）肽的毒性作用以及新型候选药物对A β肽的保护作用是一项困难的任务。广泛使用的MTT法在细胞系或原代细胞培养中可能对A β肽不敏感，本文描述了一种简单而相关的方法来测试A β 1 - 42对大鼠急性海马脑片的毒性。采用MTT法和LDH法测定不同条件下脑片的存活率。同时使用这两种测定法可以给出A β 1 - 42在缺糖缺氧（OGD）急性脑切片模型中毒性作用的详细相关结果。两种测定法都能够通过测量光密度（OD）来定量组织活力。我们发现，同时应用OGD和A β 1 - 42处理诱导海马脑片活力的降低比它们单独的作用更强烈，使用MTT和LDH测定定量脑片活力被证明是一种简单的离体方法来研究A β毒性。测试脑切片在阿尔茨海默病研究中比使用体外细胞培养物更相关，这是由于维持了三维细胞网络、细胞变异性和完整的细胞连接。(C)2012 Elsevier Inc. All rights reserved.
It is difficult task to measure precisely the toxic effect of beta-amyloid (A beta 1-42) peptides and also the protective effect of novel drug candidates against A beta-peptides. The widely used MTT-assay in cell lines or primary cell cultures could be insensitive against A beta-peptides.We describe here an easy and relevant method for testing A beta 1-42 toxicity on acute hippocampal slices derived from rat. Brain slice viability in different conditions was measured using MTT and LDH assays. The concomitant use of these two assays can give detailed and relevant results on the toxic effect of A beta 1-42 in oxygen-glucose deprived (OGD) acute brain slice model. Both assays are capable of quantifying tissue viability by measuring optical density (OD). We found that simultaneous application of OGD and A beta 1-42 treatment induced a more intensive decrease in hippocampal slice viability than their separate effects.The use of MTT and LDH assay for quantifying brain slice viability proved to be an easy ex vivo method for investigating A beta toxicity. Testing brain slices is more relevant in Alzheimer's Disease research than using in vitro cell cultures, due to maintenance of the three dimensional cellular network, the cell variability and intact cell connections. (C) 2012 Elsevier Inc. All rights reserved.