Isolation and Detection of Pathological Tau Species in a Tauopathy Mouse Model

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2761; p. 317
• Main Authors: Singh, Abhay Kumar, Selvarasu, Karthikeyan, Durairajan, Siva Sundara Kumar
• Format: Journal Article
• Language: English
• Published: United States 2024
• Subjects: Alzheimer Disease - metabolism; Animals; Brain - metabolism; Disease Models, Animal; Mice; Mice, Transgenic; tau Proteins - metabolism; Tauopathies - metabolism; Immunohistochemistry; Tauopathies; Ultracentrifugation; Insoluble phosphotau; Transgenic tau mouse model; Western blotting
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-3662-6_23

Tau protein in Alzheimer's disease (AD) and tauopathies becomes insoluble due to hyperphosphorylation, conformational alterations, and aggregation. To analyze insoluble tau and pathological tau species, this study employs a methodology that utilizes wild-type and transgenic tau mice (P310S Tau) tissue extraction using 1% Sarkosyl or N-Lauroylsarcosine sodium salt and the radio immunoprecipitation assay (RIPA) buffer. However, the commonly used methods to study the insoluble tau fraction using detergents like Sarkosyl and RIPA require a large amount of homogenate, which can pose challenges when dealing with small tissue samples. Additionally, the study employs immunohistochemistry to visualize and quantify the pathological tau species in the brain tissue of transgenic mice, aiming to identify and analyze pathological tau species such as hyperphosphorylated tau to further our understanding of tauopathies such as Alzheimer's disease.