Carbamazepine induction of apoptosis in cultured cerebellar neurons: effects ofN-methyl-d-aspartate, aurintricarboxylic acid and cycloheximide

Xiao Ming Gao, Russell Louis Margolis, Peter Leeds, Christopher Hough, Robert M. Post, De Maw Chuang

Research output: Contribution to journalArticle

Abstract

We have previously demonstrated that carbamazepine (CBZ) at concentrations above the therapeutic range is toxic to cultured cerebellar granule cells. Here, we ask whether the effect of CBZ involves neuronal apoptosis or necrosis. Treatment of cultured cerebellar granule cells with CBZ for 3 days resulted in a concentration-dependent fragmentation of DNA revealed as a laddered pattern in agarose gel electrophoresis, a phenomenon characteristic of apoptosis. Pretreatment of cells withN-methyl-d-aspartate, (NMDA) blocked CBZ-induced DNA fragmentation and neurotoxicity as assayed by loss of mitochondrial activity with MTT or by [3H]ouabain binding to Na+/K+-ATPase. Aurintricarboxylic acid (ATA), a polyanionic dye, also markedly suppressed DNA fragmentation and cell death detected by morphological examination. A considerable level of DNA ladder formation was detected in untreated cells and this basal DNA fragmentation was also blocked by NMDA and ATA. Moreover, NMDA and ATA prevented CBZ-induced chromatin condensation as revealed by DNA binding with the fluorescent dye Hoechst 33258. Pretreatment of cells with cycloheximide, a protein synthesis inhibitor, prevented CBZ-induced cell death detected morphologically and attenuated CBZ-induced neurotoxicity assessed by mitochondrial activity and [3H]ouabain binding assays. Taken together, our results suggest that CBZ-induces death of cerebellar granule cells by an apoptotic process that is sensitive to NMDA, ATA and cycloheximide.

Original languageEnglish (US)
Pages (from-to)63-71
Number of pages9
JournalBrain Research
Volume703
Issue number1-2
DOIs
StatePublished - Dec 12 1995
Externally publishedYes

Fingerprint

Aurintricarboxylic Acid
Carbamazepine
Cycloheximide
Aspartic Acid
Apoptosis
Neurons
DNA Fragmentation
N-Methylaspartate
Ouabain
Cell Death
Bisbenzimidazole
Protein Synthesis Inhibitors
Agar Gel Electrophoresis
Poisons
DNA
Fluorescent Dyes
Chromatin
Necrosis
Coloring Agents

Keywords

  • Apoptosis
  • Aurintricarboxylic acid
  • Carbamazepine
  • Cerebellar granule cell
  • Cycloheximide
  • DNA fragmentation
  • N-Methyl-d-aspartate

ASJC Scopus subject areas

  • Developmental Biology
  • Molecular Biology
  • Clinical Neurology
  • Neuroscience(all)

Cite this

Carbamazepine induction of apoptosis in cultured cerebellar neurons : effects ofN-methyl-d-aspartate, aurintricarboxylic acid and cycloheximide. / Gao, Xiao Ming; Margolis, Russell Louis; Leeds, Peter; Hough, Christopher; Post, Robert M.; Chuang, De Maw.

In: Brain Research, Vol. 703, No. 1-2, 12.12.1995, p. 63-71.

Research output: Contribution to journalArticle

Gao, Xiao Ming ; Margolis, Russell Louis ; Leeds, Peter ; Hough, Christopher ; Post, Robert M. ; Chuang, De Maw. / Carbamazepine induction of apoptosis in cultured cerebellar neurons : effects ofN-methyl-d-aspartate, aurintricarboxylic acid and cycloheximide. In: Brain Research. 1995 ; Vol. 703, No. 1-2. pp. 63-71.
@article{4d4fa08069fd4807b7ee5385407bdb9e,
title = "Carbamazepine induction of apoptosis in cultured cerebellar neurons: effects ofN-methyl-d-aspartate, aurintricarboxylic acid and cycloheximide",
abstract = "We have previously demonstrated that carbamazepine (CBZ) at concentrations above the therapeutic range is toxic to cultured cerebellar granule cells. Here, we ask whether the effect of CBZ involves neuronal apoptosis or necrosis. Treatment of cultured cerebellar granule cells with CBZ for 3 days resulted in a concentration-dependent fragmentation of DNA revealed as a laddered pattern in agarose gel electrophoresis, a phenomenon characteristic of apoptosis. Pretreatment of cells withN-methyl-d-aspartate, (NMDA) blocked CBZ-induced DNA fragmentation and neurotoxicity as assayed by loss of mitochondrial activity with MTT or by [3H]ouabain binding to Na+/K+-ATPase. Aurintricarboxylic acid (ATA), a polyanionic dye, also markedly suppressed DNA fragmentation and cell death detected by morphological examination. A considerable level of DNA ladder formation was detected in untreated cells and this basal DNA fragmentation was also blocked by NMDA and ATA. Moreover, NMDA and ATA prevented CBZ-induced chromatin condensation as revealed by DNA binding with the fluorescent dye Hoechst 33258. Pretreatment of cells with cycloheximide, a protein synthesis inhibitor, prevented CBZ-induced cell death detected morphologically and attenuated CBZ-induced neurotoxicity assessed by mitochondrial activity and [3H]ouabain binding assays. Taken together, our results suggest that CBZ-induces death of cerebellar granule cells by an apoptotic process that is sensitive to NMDA, ATA and cycloheximide.",
keywords = "Apoptosis, Aurintricarboxylic acid, Carbamazepine, Cerebellar granule cell, Cycloheximide, DNA fragmentation, N-Methyl-d-aspartate",
author = "Gao, {Xiao Ming} and Margolis, {Russell Louis} and Peter Leeds and Christopher Hough and Post, {Robert M.} and Chuang, {De Maw}",
year = "1995",
month = "12",
day = "12",
doi = "10.1016/0006-8993(95)01066-1",
language = "English (US)",
volume = "703",
pages = "63--71",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Carbamazepine induction of apoptosis in cultured cerebellar neurons

T2 - effects ofN-methyl-d-aspartate, aurintricarboxylic acid and cycloheximide

AU - Gao, Xiao Ming

AU - Margolis, Russell Louis

AU - Leeds, Peter

AU - Hough, Christopher

AU - Post, Robert M.

AU - Chuang, De Maw

PY - 1995/12/12

Y1 - 1995/12/12

N2 - We have previously demonstrated that carbamazepine (CBZ) at concentrations above the therapeutic range is toxic to cultured cerebellar granule cells. Here, we ask whether the effect of CBZ involves neuronal apoptosis or necrosis. Treatment of cultured cerebellar granule cells with CBZ for 3 days resulted in a concentration-dependent fragmentation of DNA revealed as a laddered pattern in agarose gel electrophoresis, a phenomenon characteristic of apoptosis. Pretreatment of cells withN-methyl-d-aspartate, (NMDA) blocked CBZ-induced DNA fragmentation and neurotoxicity as assayed by loss of mitochondrial activity with MTT or by [3H]ouabain binding to Na+/K+-ATPase. Aurintricarboxylic acid (ATA), a polyanionic dye, also markedly suppressed DNA fragmentation and cell death detected by morphological examination. A considerable level of DNA ladder formation was detected in untreated cells and this basal DNA fragmentation was also blocked by NMDA and ATA. Moreover, NMDA and ATA prevented CBZ-induced chromatin condensation as revealed by DNA binding with the fluorescent dye Hoechst 33258. Pretreatment of cells with cycloheximide, a protein synthesis inhibitor, prevented CBZ-induced cell death detected morphologically and attenuated CBZ-induced neurotoxicity assessed by mitochondrial activity and [3H]ouabain binding assays. Taken together, our results suggest that CBZ-induces death of cerebellar granule cells by an apoptotic process that is sensitive to NMDA, ATA and cycloheximide.

AB - We have previously demonstrated that carbamazepine (CBZ) at concentrations above the therapeutic range is toxic to cultured cerebellar granule cells. Here, we ask whether the effect of CBZ involves neuronal apoptosis or necrosis. Treatment of cultured cerebellar granule cells with CBZ for 3 days resulted in a concentration-dependent fragmentation of DNA revealed as a laddered pattern in agarose gel electrophoresis, a phenomenon characteristic of apoptosis. Pretreatment of cells withN-methyl-d-aspartate, (NMDA) blocked CBZ-induced DNA fragmentation and neurotoxicity as assayed by loss of mitochondrial activity with MTT or by [3H]ouabain binding to Na+/K+-ATPase. Aurintricarboxylic acid (ATA), a polyanionic dye, also markedly suppressed DNA fragmentation and cell death detected by morphological examination. A considerable level of DNA ladder formation was detected in untreated cells and this basal DNA fragmentation was also blocked by NMDA and ATA. Moreover, NMDA and ATA prevented CBZ-induced chromatin condensation as revealed by DNA binding with the fluorescent dye Hoechst 33258. Pretreatment of cells with cycloheximide, a protein synthesis inhibitor, prevented CBZ-induced cell death detected morphologically and attenuated CBZ-induced neurotoxicity assessed by mitochondrial activity and [3H]ouabain binding assays. Taken together, our results suggest that CBZ-induces death of cerebellar granule cells by an apoptotic process that is sensitive to NMDA, ATA and cycloheximide.

KW - Apoptosis

KW - Aurintricarboxylic acid

KW - Carbamazepine

KW - Cerebellar granule cell

KW - Cycloheximide

KW - DNA fragmentation

KW - N-Methyl-d-aspartate

UR - http://www.scopus.com/inward/record.url?scp=0028879386&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028879386&partnerID=8YFLogxK

U2 - 10.1016/0006-8993(95)01066-1

DO - 10.1016/0006-8993(95)01066-1

M3 - Article

C2 - 8719616

AN - SCOPUS:0028879386

VL - 703

SP - 63

EP - 71

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1-2

ER -