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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
RÉSUMÉ

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).

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Hydroxyde de sodium, ACS reagent, ≥97.0%, pellets
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Acide acétique, glacial, ReagentPlus®, ≥99%
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Solution tampon phosphate, powder, pH 7.4, for preparing 1 L solutions
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DAPI, for nucleic acid staining
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Cresyl Violet acetate, certified by the Biological Stain Commission
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TWEEN® 20, viscous liquid
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Polyvinylpyrrolidone, average mol wt 40,000
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Anticorps anti-protéine NeuN purifiée, from guinea pig, purified by affinity chromatography
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Anticorps anti-caspase-3, forme active (clivée), Chemicon®, from rabbit