Korean J Physiol Pharmacol.
1997 Dec;1(6):699-705.
Apoptotic process is involved in the L-glutamate-induced PC12 cell
death
- Affiliations
-
- 1
Department of Pharmacology, Catholic University Medical College, Seoul 137-701, South Korea.
Abstract
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Although it is known that neuronal cell death during development occurs
by apoptosis, the mechanisms underlying excitatory amino acid-induced
neuronal cell death remain poorly understood. In this study we have
examined the mechanism by which L-glutamate, an excitatory amino acid
neurotransmitter, induces cell death in PC12 cell lines. To
characterize cell death, we employed sandwich enzyme-linked
immunosorbent assay (ELISA) method for cellular DNA fragmentation, DNA
agarose gel electrophoresis and chromatin staining by acridine orange
and ethidium bromide after treating the PC12 cells with L-glutamate.
L-Glutamate caused dose-dependent cell death with a maximum at 24 hrs
after the treatment. These cellular fragmentation was blocked by
pretreatment of MK-801, a noncompetitive N-methyl-D-aspartic acid
(NMDA) receptor antagonist, and nerve growth factor(NGF). Analysis of
DNA integrity from L-glutamate-treated cells revealed cleavage of DNA
into regular sized fragments, a biochemical hallmark of apoptosis. The
PC12 cells that were induced to die by L-glutamate treatment exhibited
classical chromatin condensation under the light microscopy after
acridine orange and ethidium bromide staining. These results suggest
that apoptosis is one of the key features that are involved in
L-glutamate-induced excitotoxic cell death in PC12 cells, and these
cell death are mediated by NMDA receptor and depend on NGF.