A novel method for the rapid determination of beta-amyloid toxicity on acute hippocampal slices using MTT and LDH assays

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• 作者：Emese Mozes^a ; ^{mese859@gmail.com} ; Akos Hunya^a ; ¹ ; ^{hunya@dnt.u-szeged.hu} ; Aniko Posa^b ; ² ; ^{paniko@bio.u-szeged.hu} ; Botond Penke^a ; ³ ; ^{pbotond@mdche.szote.u-szeged.hu} ; Zsolt Datki^a ; ⁴ ; ^{datkiz@yahoo.com}
• 关键词：ACSF ; artificial cerebrospinal fluid ; AD ; Alzheimer's disease ; A尾 ; amyloid-beta ; HEPES ; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ; LDH ; Lactate dehydrogenase ; MTT ; 3-(4 ; 5-dimethylthiazol-2-yl)-2 ; 5-diphenyltetrazolium bromide ; OD ; optical density
• 刊名：Brain Research Bulletin
• 出版年：2012
• 期刊代码：9_03619230
• 类别：neu
• 出版时间：10 April, 2012
• 卷：87
• 期：6
• 页码：521-525
• 文件大小：489 K

摘要

It is difficult task to measure precisely the toxic effect of beta-amyloid (A尾 1-42) peptides and also the protective effect of novel drug candidates against A尾-peptides. The widely used MTT-assay in cell lines or primary cell cultures could be insensitive against A尾-peptides.

We describe here an easy and relevant method for testing A尾 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尾 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尾 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尾 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.