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Assessment of neurodegeneration and neuronal loss in aged 5XFAD mice.

STAR protocols (2021-11-11)
Liansheng Zhang, Jie Li, Anning Lin
ABSTRACT

Neuronal loss resulting from progressive neurodegeneration is a major pathological feature of Alzheimer's disease (AD). Here, we present a protocol to detect neurodegeneration, neuronal apoptosis, and neuronal loss in 5XFAD mouse strain, which is a well-established model for interrogating the molecular mechanism of neuronal death in AD. This protocol describes the use of the neurodegenerative marker Fluro-Jade C, cleaved caspase-3 immunofluorescent staining and Nissl staining for the analysis of neurodegeneration and neuronal loss in 5XFAD mice. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
TWEEN® 20, viscous liquid
Sigma-Aldrich
Cresyl Violet acetate, certified by the Biological Stain Commission
Sigma-Aldrich
Anti-Caspase 3 Antibody, active (cleaved) form, Chemicon®, from rabbit
Sigma-Aldrich
Sodium hydroxide, ACS reagent, ≥97.0%, pellets
Sigma-Aldrich
Acetic acid, glacial, ReagentPlus®, ≥99%
Sigma-Aldrich
Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Polyvinylpyrrolidone, average mol wt 40,000
Sigma-Aldrich
Anti-NeuN purified Antibody, from guinea pig, purified by affinity chromatography