Increased cholesterol in Abeta-positive nerve terminals from Alzheimer's disease cortex.
Synapse loss in Alzheimer's disease (AD) is poorly understood but evidence suggests it is a key pathological event. In order to precisely detect stable synaptic changes, we have developed methods for flow cytometry analysis of synaptosomes prepared from cryopreserved AD samples, and have previously shown that amyloid-beta (Abeta) accumulates in surviving presynaptic terminals in AD cortex. In the present experiments we have examined amyloid-containing terminals in more detail, first dual labeling synaptosomes from AD cortex for Abeta and a series of markers, and then using quadrant analysis to compare amyloid-positive and amyloid-negative terminals. Amyloid-positive synaptosomes were larger in size than amyloid-negatives (p<0.007), and significant increases were observed in mean fluorescence for the lipid raft markers cholesterol (27%; p<0.0005) and GM1 ganglioside (24%; p<0.005). SNAP-25 immunofluorescence was increased by 31% (p<0.0001) in amyloid-bearing terminals, consistent with a sprouting response to amyloid accumulation. These results suggest that Abeta accumulation in synaptic terminals may underly dysfunction prior to or independent of extracellular amyloid deposition.
Gylys
Karen Hoppens
KH
UCLA School of Nursing, Los Angeles, CA 90095, USA. kgylys@sonnet.ucla.edu
Fein
Jeffrey A
JA
Yang
Fusheng
F
Miller
Carol A
CA
Cole
Gregory M
GM
D000222:Adaptation, Physiological; D000368:Aged; D000544:Alzheimer Disease / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism; D019579:Neocortex / Q000378:metabolism*; D049128:Sphingolipid Activator Proteins / Q000378:metabolism*; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism*; D013574:Synaptosomes / Q000378:metabolism*
D016229:Amyloid beta-Peptides; C494208:SNAP25 protein, human; D049128:Sphingolipid Activator Proteins; D050825:Synaptosomal-Associated Protein 25; D002784:Cholesterol