GM1-ganglioside-induced Abeta assembly on synaptic membranes of cultured neurons.
The cell-surface expression of GM1 ganglioside was studied using various cultured cells, including brain-derived endothelial cells, astrocytes, neuroblastoma cells (SH-SY5Y), and pheochromocytoma cells (PC12). GM1 ganglioside was detected only on the surface of native and nerve-growth-factor (NGF)-treated PC12 cells. We investigated whether GM1 ganglioside on the surface of these cells is sufficiently potent to induce the assembly of an exogenous soluble amyloid beta-protein (Abeta). A marked Abeta assembly was observed in the culture of NGF-treated PC12 cells. Notably, immunocytochemical study revealed that, despite the ubiquitous surface expression of GM1 ganglioside throughout cell bodies and neurites, Abeta assembly initially occurred at the terminals of SNAP25-immunopositive neurites. Abeta assembly in the culture was completely suppressed by the coincubation of Abeta with the subunit B of cholera toxin, a natural ligand for GM1 ganglioside, or 4396C, a monoclonal antibody specific to GM1-ganglioside-bound Abeta (GAbeta). In primary neuronal cultures, Abeta assembly initially occurred at synaptophysin-positive sites. These results suggest that the cell-surface expression of GM1 ganglioside is strictly cell-type-specific, and that expression of GM1 ganglioside on synaptic membranes is unique in terms of its high potency to induce Abeta assembly through the generation of GAbeta, which is an endogenous seed for Abeta assembly in Alzheimer brain.
Yamamoto
Naoki
N
Department of Alzheimer's Disease Research National Institute for Longevity Sciences National Center for Geriatrics and Gerontology 36-3 Gengo, Morioka, Obu 474-8522, Japan.
Fukata
Yuko
Y
Fukata
Masaki
M
Yanagisawa
Katsuhiko
K
D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D005677:G(M1) Ganglioside / Q000378:metabolism*; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D020932:Nerve Growth Factor / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016716:PC12 Cells; D017729:Presynaptic Terminals / Q000187:drug effects / Q000378:metabolism; D020836:Protein Structure, Quaternary; D051381:Rats; D013570:Synaptic Membranes / Q000187:drug effects / Q000378:metabolism*
D016229:Amyloid beta-Peptides; D009025:Morpholines; D005677:G(M1) Ganglioside; C033110:RV 538; D002772:Cholera Toxin; D020932:Nerve Growth Factor