Glutamate-induced antigenic changes of phospholipase C-delta in cultured cortical neurons.
Phosphoinositide-specific phospholipase C (PLC) is a key enzyme in signal transduction. It was previously demonstrated that an antibody to an isozyme of PLC, PLC-delta, produces intense staining of neurofibrillary tangles (NFT), the neurites surrounding senile plaque (SP) cores and neuropil threads in the brains of patients with Alzheimer's disease (AD). Although the etiology of neuronal degeneration in AD is still to be defined, excitotoxic glutamate might be a candidate. In the present study, an anti-PLC-delta antibody was used to examine the influence of glutamate on PLC-delta immunoreactivity in cultured rat cortical neurons. Exposure to glutamate caused the death of cultured cortical neurons and exhibited increased immunostaining with the anti-PLC-delta antibody. Subtoxic doses of glutamate also increased PLC-delta immunoreactivity in a dose-dependent manner. Both glutamate-induced neuronal degeneration and the increases in PLC-delta immunoreactivity were prevented by removal of extracellular Ca2+ or the application of an N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801. The glutamate-induced increase in PLC-delta immunoreactivity was also prevented by N omega-nitro-L-arginine, a nitric oxide (NO) synthase inhibitor. These results suggest that NO formation secondary to Ca2+ influx by NMDA receptor activation leads to similar modifications of PLC-delta to those seen in AD.
Shimohama
S
S
Department of Neurology, Faculty of Medicine, Kyoto University, Japan.
Akaike
A
A
Tamura
Y
Y
Matsushima
H
H
Kume
T
T
Fujimoto
S
S
Takenawa
T
T
Kimura
J
J
D000818:Animals; D000918:Antibody Specificity; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D002540:Cerebral Cortex / Q000166:cytology*; D018698:Glutamic Acid / Q000494:pharmacology*; D007527:Isoenzymes / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D009498:Neurotoxins / Q000494:pharmacology; D054803:Phospholipase C delta; D051381:Rats; D010738:Type C Phospholipases / Q000187:drug effects* / Q000276:immunology
D007527:Isoenzymes; D009498:Neurotoxins; D018698:Glutamic Acid; D010738:Type C Phospholipases; D054803:Phospholipase C delta