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

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.

Keyword

Glutamate; Excitotoxicity; PCl2 cell; Apoptosis; NMDA; NGF

MeSH Terms

Acridine Orange
Animals
Apoptosis
Cell Death
Chromatin
Dizocilpine Maleate
DNA
DNA Fragmentation
Electrophoresis, Agar Gel
Enzyme-Linked Immunosorbent Assay
Ethidium
Excitatory Amino Acids
Glutamic Acid
Microscopy
N-Methylaspartate
Nerve Growth Factor
Neurons
Neurotransmitter Agents
PC12 Cells*
Acridine Orange
Chromatin
DNA
Dizocilpine Maleate
Ethidium
Excitatory Amino Acids
Glutamic Acid
N-Methylaspartate
Nerve Growth Factor
Neurotransmitter Agents
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