Document Type
Article
Publication Date
5-19-2022
Abstract
Extracellular accumulation of amyloid-beta (Aβ) plaques is one of the major pathological hallmarks of Alzheimer's disease (AD), and is the target of the only FDA-approved disease-modifying treatment for AD. Accordingly, the use of transgenic mouse models that overexpress the amyloid precursor protein and thereby accumulate cerebral Aβ plaques are widely used to model human AD in mice. Therefore, immunoassays, including enzyme-linked immunosorbent assay (ELISA) and immunostaining, commonly measure the Aβ load in brain tissues derived from AD transgenic mice. Though the methods for Aβ detection and quantification have been well established and documented, the impact of the size of the region of interest selected in the brain tissue on Aβ load measurements following immunostaining has not been reported. Therefore, the current protocol aimed to compare the Aβ load measurements across the full- and sub-regions of interest using an image analysis software. The steps involved in brain tissue preparation, free-floating brain section immunostaining, imaging, and quantification of Aβ load in full- versus sub-regions of interest are described using brain sections derived from 13-month-old APP/PS1 double transgenic male mice. The current protocol and the results provide valuable information about the impact of the size of the region of interest on Aβ-positive area quantification, and show a strong correlation between the Aβ-positive area obtained using the full- and sub-regions of interest analyses for brain sections derived from 13-month-old male APP/PS1 mice that show widespread Aβ deposition.
Recommended Citation
Ohno, Y., Murphy, R., Choi, M., Ou, W., Sumbria, R.K. Full- versus Sub-Regional Quantification of Amyloid-Beta Load on Mouse Brain Sections. J. Vis. Exp. (183), e63669, https://doi.org/10.3791/63669 (2022).
Copyright
JoVE
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Included in
Amino Acids, Peptides, and Proteins Commons, Animal Experimentation and Research Commons, Animals Commons, Medical Neurobiology Commons, Medicinal and Pharmaceutical Chemistry Commons, Nervous System Diseases Commons, Neurosciences Commons, Other Chemicals and Drugs Commons, Pharmaceutical Preparations Commons, Pharmaceutics and Drug Design Commons
Comments
This article was originally published in Journal of Visualized Experiments (JoVE), volume 183, in 2022. https://doi.org/10.3791/63669