Influence of resveratrol on the sorption of phosphorylated tau protein and MAPT on the PVDF membrane in a mouse model of Alzheimer’s disease
Abstract
Alzheimer’s disease is a complex neurodegenerative disease based on various processes associated with the accumulation and aggregation of defective proteins. Among them, the particularly important ones are the following: amyloid-β, which is formed by the breakdown of amyloid precursor protein, the accumulation of hyperphosphorylated tau proteins inside neurons that form neurofibrillary tangles, and aberrant aggregation and inclusion formation of microtubule-associated protein tau (MAPT). APP/PS1 transgenic mice act as a model of Alzheimer’s disease and express mutant human genes that cause the accumulation of amyloid-β peptides in the brain. The goal of this work was a quantitative assessment of the level of p-tau231 in the brain of transgenic mice with a model of AD using the sorption method. The objectives of the study also included testing the ability of the natural polyphenol resveratrol to reduce the concentration of p-tau231 in the brain of transgenic mice and improve their cognitive functions. Western blot is a widely used method for the immunodetection and in vitro quantitative determination of proteins.
Western blot allows separating proteins based on their molecular weight with the further transfer to an adsorption membrane. In this case, the proteins are transferred from the gel to the PVDF membrane using electrophoretic elution. This method involves placing a protein-containing polyacrylamide gel in direct contact with a PVDF membrane represented by a linear polymer with repetitive links -(CF2-CH2)-. Proteins transferred to the membrane are well-retained on its surface during the whole immunodetection process due to a combination of dipole and hydrophobic interactions. Western blot showed that mice with impaired protein aggregation accumulated significantly more MAPT and phosphorylated tau protein in the brain as compared to wild mice. In addition, in the course of the Morris water maze test, these mice showed cognitive deficits, which manifested both in the difficulty of finding the platform and more anxious behaviour, which confirmed pronounced thigmotaxis. The natural polyphenol resveratrol partially reversed cognitive deficits, although this effect was not associated with decreased levels of phosphorylated tau and MAPT.
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