Rapid detection of protein aggregates in the brains of Alzheimer patients and transgenic mouse models of amyloidosis.
Extracellular and/or intracellular aggregates are pathological features of many, if not all, neurodegenerative diseases. In Alzheimer disease (AD), extracellular aggregates of beta-amyloid (Abeta) and intracellular aggregates of tau or a-synuclein are key diagnostic markers of the disease. We report here a method to rapidly detect these protein aggregates that relies on size exclusion filtration and immunostaining of trapped material, a method termed filter trapping. We demonstrate that aggregated forms of Abeta and tau are readily trapped in 0.22 microm cellulose acetate filter membranes, which can then be immunostained with specific antibodies in a manner similar to the standard immunoblot. Coupling this method with serial dilution permits a rapid assessment of relative aggregate burden.
Xu
Guilian
G
Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Gonzales
Victoria
V
Borchelt
David R
DR
D000544:Alzheimer Disease / Q000473:pathology*; D016229:Amyloid beta-Peptides / Q000032:analysis*; D000686:Amyloidosis / Q000473:pathology*; D000818:Animals; D001921:Brain / Q000473:pathology*; D002482:Cellulose / Q000031:analogs & derivatives*; D005110:Extracellular Space / Q000000981:diagnostic imaging; D005374:Filtration; D006801:Humans; D007424:Intracellular Fluid / Q000000981:diagnostic imaging; D016631:Lewy Bodies / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000032:analysis*; D051843:Synucleins; D014463:Ultrasonography; D016875:tau Proteins / Q000032:analysis*
D016229:Amyloid beta-Peptides; D009419:Nerve Tissue Proteins; D051843:Synucleins; D016875:tau Proteins; C005062:acetylcellulose; D002482:Cellulose