Bharadwaj Prashant, Martins Ralph
Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Edith Cowan University; School of Pharmacy and Biomedical Sciences, Curtin Health and Innovation Research Institute (CHIRI), Faculty of Health Sciences, Curtin University, Western Australia, Australia.
Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Edith Cowan University; School of Psychiatry and Clinical Neuroscience, University of Western Australia, Western Australia; School of Biomedical Science, Macquarie University, Sydney, NSW, Australia.
Neural Regen Res. 2020 Oct;15(10):1931-1936. doi: 10.4103/1673-5374.280318.
Multiple lines of evidence show that soluble oligomer forms of amyloid β protein (Aβ) 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 Aβ 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 Aβ 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 Aβ induced cell death in mammalian and yeast cells. This assay offers a platform for studying underlying mechanisms of oligomer Aβ induced cell death using gene deletion/overexpression libraries and developing novel agents that alleviate Aβ induced cell death.
多条证据表明，淀粉样β蛋白（Aβ）的可溶性寡聚体形式是大脑中最具神经毒性的物质，且与阿尔茨海默病中神经元丧失的程度和认知缺陷相关。尽管许多研究已使用哺乳动物细胞来研究寡聚体Aβ的毒性，但使用更简单的真核细胞系统对大规模筛选研究具有优势。我们之前已建立并验证出芽酵母酿酒酵母是研究Aβ毒性的简单且强大的模型。使用基于菌落计数的方法，显示寡聚体Aβ可在酵母中诱导剂量依赖性细胞死亡。我们通过开发基于吸光度的生长测定法，将该方法应用于高通量筛选。我们用先前已证明可保护哺乳动物和酵母细胞中寡聚体Aβ诱导的细胞死亡的处理方法进一步验证了该测定法。该测定法提供了一个平台，可用于使用基因缺失/过表达文库研究寡聚体Aβ诱导细胞死亡的潜在机制，并开发减轻Aβ诱导细胞死亡的新型药物。
