A rapid absorbance-based growth assay to screen the toxicity of oligomer Aβ42 and protect against cell death in yeast  

作　　者：Prashant Bharadwaj Ralph Martins 

机构地区：[1]Centre of Excellence for Alzheimer’s Disease Research and Care,School of Medical and Health Sciences,Edith Cowan University,Western Australia,Australia [2]School of Pharmacy and Biomedical Sciences,Curtin Health and Innovation Research Institute(CHIRI),Faculty of Health Sciences,Curtin University,Western Australia,Australia [3]School of Psychiatry and Clinical Neuroscience.University of Western Australia,Western Australia,Australia [4]School of Biomedical Science,Macquarie University,Sydney,NSW,Australia

出　　处：《Neural Regeneration Research》2020年第10期1931-1936,共6页中国神经再生研究（英文版）

基　　金：supported by the National Health and Medical Research Council-Australian Research Council dementia research development fellowship(APP1107109)to PB

摘　　要：Multiple lines of evidence show that soluble oligomer forms of amyloidβprotein(Aβ42)are the most neurotoxic species in the brain and correlates with the degree of neuronal loss and cognitive deficit in Alzheimer’s disease.Although many studies have used mammalian cells to investigate oligomer Aβ42 toxicity,the use of more simple eukaryotic cellular systems offers advantages for large-scale screening studies.We have previously established and validated budding yeast,Saccharomyces cerevisiae to be a simple and a robust model to study the toxicity of Aβ.Using colony counting based methods,oligomeric Aβ42 was shown to induce dose-dependent cell death in yeast.We have adapted this method for high throughput screening by developing an absorbance-based growth assay.We further validated the assay with treatments previously shown to protect oligomer Aβ42 induced cell death in mammalian and yeast cells.This assay offers a platform for studying underlying mechanisms of oligomer Aβ42 induced cell death using gene deletion/overexpression libraries and developing novel agents that alleviate Aβ42 induced cell death.Multiple lines of evidence show that soluble oligomer forms of amyloid β protein(Aβ42) are the most neurotoxic species in the brain and correlates with the degree of neuronal loss and cognitive deficit in Alzheimer’s disease. Although many studies have used mammalian cells to investigate oligomer Aβ42 toxicity, the use of more simple eukaryotic cellular systems offers advantages for large-scale screening studies. We have previously established and validated budding yeast, Saccharomyces cerevisiae to be a simple and a robust model to study the toxicity of Aβ. Using colony counting based methods, oligomeric Aβ42 was shown to induce dose-dependent cell death in yeast. We have adapted this method for high throughput screening by developing an absorbance-based growth assay. We further validated the assay with treatments previously shown to protect oligomer Aβ42 induced cell death in mammalian and yeast cells. This assay offers a platform for studying underlying mechanisms of oligomer Aβ42 induced cell death using gene deletion/overexpression libraries and developing novel agents that alleviate Aβ42 induced cell death.

关 键 词：Alzheimer’s disease amyloid toxicity autophagy Aβ42 oligomer high-throughput screening latrepirdine NEUROPROTECTION yeast model 

分 类 号：R446[医药卫生—诊断学] R364[医药卫生—临床医学]