Flow cytometric analysis on cytotoxic action of amyloid beta protein fragment 25-35 on brain neurons dissociated from the rats.
Effects of amyloid beta protein fragment 25-35, A beta P(25-35), on membrane permeability and cell viability were examined in the brain neurons dissociated from the rats using a flow cytometer and two fluorescent dyes, fluo-3 to monitor intracellular Ca2+ concentration ([Ca2+]i) of neurons and ethidium which is impermeant to membranes of intact neurons to stain dead and dying neurons. A beta P(25-35) augmented fluo-3 fluorescence of some neurons at concentrations greater than 1 microM, indicating an increase in [Ca2+]i although other neurons (about 80% of total neurons) did not respond to A beta P(25-35) even at 10 microM. A beta P(25-35) at 1 microM or greater increased dose-dependently the number of ethidium-stained neurons, suggesting a dose-dependent increase in number of dead and dying neurons. Results suggest that A beta P(25-35) increases the membrane permeability of brain neurons, resulting in a destabilized intracellular homeostasis that leads to neuonal death.
Laboratory of Cell Signaling [Pharmacology], Faculty of Integrated Arts and Sciences, University of Tokushima, Japan.
K
K
Furukawa
Y
Y
Oyama
L
L
Chikahisa
Y
Y
Hatakeyama
N
N
Akaike
D016229:Amyloid beta-Peptides / Q000633:toxicity*; D000814:Aniline Compounds; D000818:Animals; D001921:Brain / Q000166:cytology* / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D004996:Ethidium; D005434:Flow Cytometry; D009474:Neurons / Q000187:drug effects*; D010446:Peptide Fragments / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D014966:Xanthenes
D016229:Amyloid beta-Peptides; D000814:Aniline Compounds; D010446:Peptide Fragments; D014966:Xanthenes; C074636:amyloid beta-protein (25-35); C059715:Fluo-3; D004996:Ethidium; D002118:Calcium