title: Triple immunofluorolabeling with two rabbit polyclonal antibodies and a mouse monoclonal antibody allowing three-dimensional analysis of cotton wool plaques in Alzheimer disease.
abstract: We established a triple-labeling method with two rabbit polyclonal antibodies and a mouse monoclonal antibody and examined autopsied brain tissue with cotton wool plaques (CWPs). One of the polyclonal antibodies was so diluted (anti-Abeta42 or anti-Abeta40/1:30,000 or anti-von Willebrand factor/1:1000) that its visualization was possible only after amplification with the catalyzed reporter deposition (CARD) method. The other polyclonal antibody (anti-Abeta40 or anti-Abeta42/1:1000) was visualized with a fluorochrome conjugated to an anti-rabbit antibody that specifically visualized the latter polyclonal antibody because of its lower sensitivity. A monoclonal antibody, AT8, was superimposed to yield triple immunofluorolabeling. Serial optical sections with an interval of 0.3 micro m were reconstructed to allow three-dimensional (3D) observation of these three epitopes. Abeta40 was localized to core-like structures, mainly in layers I-III, and was sometimes in contact with the vascular wall, both without neuritic reactions. CWPs, present in layers I-VI, were labeled with anti-Abeta42 and were accompanied by neuritic reactions. These differences suggest that mechanisms of Abeta deposition and its relation to neuritic reactions or to blood vessels differ according to the lesion, even in the same microscopic field.
authors.affiliation: Departments of Neuropathology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan. uchihara@tmin.ac.jp
authors.forename: Toshiki
authors.initials: T
authors.lastname: Uchihara
authors.forename: Ayako
authors.initials: A
authors.lastname: Nakamura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakayama
authors.forename: Kunimasa
authors.initials: K
authors.lastname: Arima
authors.forename: Norio
authors.initials: N
authors.lastname: Ishizuka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mori
authors.forename: Setsuo
authors.initials: S
authors.lastname: Mizushima
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology*; D016229:Amyloid beta-Peptides / Q000276:immunology / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D018613:Microscopy, Confocal; D016874:Neurofibrillary Tangles / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D058225:Plaque, Amyloid / Q000378:metabolism*; D011817:Rabbits; D016875:tau Proteins / Q000276:immunology / Q000378:metabolism; D014841:von Willebrand Factor / Q000276:immunology / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; C078592:amyloid beta-protein (1-40); C075222:amyloid beta-protein (1-42); D016875:tau Proteins; D014841:von Willebrand Factor