Assessing β-amyloid-induced NLRP3 inflammasome activation in primary microglia.

Senile plaques are an important histological hallmark of Alzheimer's disease. They mainly consist of the fibrillar peptide β-amyloid (Aβ) and are surrounded by activated microglia and astrocytes. Microglia in the vicinity of senile plaques express high levels of proinflammatory cytokines and neurotoxic substances, which are believed to influence disease progression. One important cytokine in Alzheimer's disease is IL-1β. Stimulation of cultured primary microglia by synthetic fibrillar Aβ causes the release of IL-1β via activation of the NLRP3 inflammasome.Here we provide protocols for the preparation of primary microglial cultures and synthetic oligomeric and fibrillar forms of Aβ.