title: Flow Cytometry Analysis and Quantitative Characterization of Tau in Synaptosomes from Alzheimer's Disease Brains.
abstract: Synaptosomes, resealed nerve terminals that form when tissue is homogenized in isotonic medium, are a model system that has been a key source of knowledge about neurotransmission. Synaptosomes contain mitochondria, cytoskeletal proteins, and release neurotransmitters; many have postsynaptic elements. Cryopreservation at the time of autopsy makes it possible to prepare synaptosomes from human samples. Flow cytometry is a powerful analytic technique that precisely measures fluorescence on a cell-by-cell basis, and also indicates particle size and complexity with a routine parameter that measures light scattering. We describe here a procedure for flow cytometry analysis of tau in synaptosomes, a procedure that enables (1) "purification" of synaptosomes from the P-2 fraction (crude synaptosomes) by gating on particle size, and (2) quantitative measure of tau immunofluorescence in individual terminals. Application of flow cytometry to study of synaptosomes has yielded important information, not possible with routine biochemistry, about synaptic pathology in Alzheimer's disease.
authors.affiliation: UCLA School of Nursing and Mary S. Easton Center for Alzheimer's Research at UCLA, Box 956919, Factor Building, Los Angeles, CA, 90095-6919, USA. kgylys@sonnet.ucla.edu.
authors.forename: Karen Hoppens
authors.initials: KH
authors.lastname: Gylys
authors.affiliation: UCLA School of Nursing and Mary S. Easton Center for Alzheimer's Research at UCLA, Box 956919, Factor Building, Los Angeles, CA, 90095-6919, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Bilousova
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism*; D000818:Animals; D015925:Cryopreservation; D003599:Cytoskeleton / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D013574:Synaptosomes / Q000378:metabolism*; D016875:tau Proteins / Q000032:analysis* / Q000378:metabolism*
chemical_list: D016875:tau Proteins
keywords: Cytoskeleton; FACs; Flow cytometry; HT7 antibody; Neurodegeneration; Tau fragment