A rapid method to measure beta-amyloid induced neurotoxicity in vitro

作者：Patel Dhara; Good Theresa^*

来源：Journal of Neuroscience Methods, 2007, 161(1): 1-10.

DOI：10.1016/j.jneymeth.2006.10.004

• Alzheimer's disease
• beta-amyloid
• SH-SY5Y
• flow cytometry
• annexin
• 7-aminoactinomycin D

beta-Amyloid (A beta) is the primary protein component of senile plaques in Alzheimer's disease (AD) and is believed to be associated with neurotoxicity in the disease. A beta-induced neurotoxicity is strongly dependent on its structure. The aggregation of the peptide from monomeric form to fibrils is a function of many variables including time. It is because of this dynamic nature of A beta structure that there is a necessity for an in vitro toxicity assay that is rapid enough to eliminate or at least lower the possibility of A beta structural changes during the time required for the assay. Here we describe a fast and sensitive method with which to assess A beta-induced neurotoxicity in SH-SY5Y neuroblastoma cells. The method employs two-color flow cytometry using annexin-phycoerythrin, a marker for cell surface phosphatidylserine that typically indicates early stages of apoptosis, and 7-amino-actinomycin D, a membrane impermeant nucleic acid dye. We compare results using the two-color assay to those obtained using the propidium iodide toxicity assay and demonstrate comparability of results, but in 2 h or less with the two-color assay as opposed to 24-48 h with the propidium iodide assay. The assay described could be a useful tool in evaluating the role of A beta structure in biological activity of the peptide.

• 出版日期2007-3-30