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Vol. 53, Issue 1, 33-42, January 1998
B Contributes to Excitotoxin-Induced Apoptosis
in Rat Striatum
Experimental Therapeutics Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892
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Summary |
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Top
Summary
Introduction
Materials & Methods
Results
Discussion
References
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Excitotoxin-induced destruction of striatal neurons, proposed as a
model of Huntington's disease, involves a process having the
biochemical stigmata of apoptosis. Recent studies suggested that
transcription factor nuclear factor (NF)-
B may be involved in
excitotoxicity. To further analyze the contribution of NFB to
excitotoxic neuronal death in vivo, changes in binding
activities of NFB and other transcription factors as well as the
consequences of inhibiting NFB nuclear translocation were measured
after the infusion of quinolinic acid (120 nmol) into rat striatum.
Internucleosomal DNA fragmentation and terminal transferase-mediated
dUTP digoxigenin nick end labeling-positive nuclei appeared 12 hr later
and intensified over the next 12 hr. NFB binding activity increased
severalfold from 2 to 12 hr, then gradually declined during the next 12 hr. Other transcription factor changes included AP-1, whose binding peaked about 6 hr after quinolinic acid administration, and E2F-1, which was only modestly and transiently elevated. In contrast, quinolinic acid lead to a reduction in OCT-1, beginning after 12 hr,
and briefly in SP-1 binding. The NFB, AP-1, and OCT-1 changes were
attenuated both by the N-methyl-D-aspartate
receptor antagonist MK-801 and the protein synthesis inhibitor
cycloheximide. Moreover, quinolinic acid-induced internucleosomal DNA
fragmentation and striatal cell death were significantly reduced by the
intrastriatal administration of NFB SN50, a cell-permeable
recombinant peptide that blocks NFB nuclear translocation. These
results illustrate the complex temporal pattern of transcription factor
change attending the apoptotic destruction produced in rat striatum by
quinolinic acid. They further suggest that NFB activation
contributes to the excitotoxin-induced death of striatal neurons.
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Introduction |
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Top
Summary
Introduction
Materials & Methods
Results
Discussion
References
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Neurodegenerative
disorders, such as HD, are characterized by the progressive loss of
specific central neurons during adult life. Although HD is caused by a
polyglutamine expansion in the gene for HD, the exact process by which
this abnormality triggers the death of striatal neurons is not yet
known. Recent studies of postmortem tissue suggest the degenerative
process may involve an apoptotic mechanism (Portera-Cailliau et
al., 1995
).
Glutamate, which can induce oxidative stress in neuronal tissue, has
been implicated in the pathogenesis of HD and other neurodegenerative disorders. Indeed, the demise of striatal neurons produced by the
glutamate receptor agonist QA as well by kainic acid has been proposed
as an animal model of HD (Coyle and Schwarcz, 1976). Recent
observations suggest that the QA- or kainic acid -induced destruction
of striatal cells occurs, at least in part, by an apoptotic mechanism
(Ankarcrona et al., 1993; Bonfoco et al., 1995;
Filipkowski et al., 1995; Gillardon et al., 1995;
Portera-Cailliau et al., 1995; Qin et al., 1996;
Simonian et al., 1996). Although the morphological features
of the apoptotic process have been well described, just how
glutamatergic receptor agonists activate cell death programs at the
molecular level remains to be elucidated.
Transcription factors, including immediate early genes such as
c-jun, E2F-1, OCT-1 and NFB, have been increasingly
implicated in the control of apoptosis (Dragunow and Preston, 1995;
Grilli et al., 1996; Ham et al., 1995; Wang and
Pittman, 1993). Induction of these regulators of gene expression
typically precedes the appearance of internucleosomal DNA fragmentation
(Estus et al., 1994). Moreover, antisense, antibody, gene
mutation, and pharmacological techniques that selectively inhibit
certain transcription factors have been found to block the death of
cultured cells, thus suggesting that these factors may be direct
contributors to the generation of apoptotic cascades (Estus et
al., 1994; Ham et al., 1995; Lin et al.,
1995a).
NFB, a member of the Rel transcription factor family,
participates in the regulation of a broad array of genes primarily involved in immune and stress defense mechanisms. It has also been
linked to the generation of certain cancers and to the control of the
cell cycle. In the central nervous system, NFB is constitutively expressed in both neurons and glia (Kaltschmidt et al.,
1994). A variety of pathogenetic stimuli, including oxidative stress, ischemic insult, and 
-amyloid deposition (Kaltschmidt et
al., 1997; Legrand-Poels et al., 1995; Salminen
et al., 1995), can release NFB from cytosolic
sequestration sites where it is bound to a member of the inhibitory
protein family, IB (Beg and Baltimore, 1996; Brown et
al., 1993; Liou and Baltimore, 1993). Upon translocated to the
nucleus, NFB acts as a positive regulator of genes favoring either
protective or degenerative responses, depending on genetic programs
within a particular cell type (Baeuerle, 1991; Baichwal and Baeuerle,
1997; Lipton, 1997). Several NFB target genes, including P53 and
c-Myc, are well established modulators of apoptosis (Wu and Lozano,
1994).
Recent in vitro studies have found that stimulation of
glutamate receptors strongly activates NFB (Guerrini et
al., 1995; Kaltschmidt et al., 1995). Subsequent
reports that NFB inhibitors such as aspirin and salicylate protect
cultured neurons against glutamate-induced neuronal toxicity (Grilli
et al., 1996) could thus indicate that NFB activation
contributes to excitotoxic neuronal injury. Unfortunately,
interpretation of these results is complicated by the fact that
salicylates inhibit other transcription factors and several protein
kinases in addition to having many other pharmacologic actions (Frantz
and O'Neill, 1995).
To more precisely delineate the role of NFB in excitotoxin-induced
neuronal apoptosis in vivo, we have examined the temporal pattern of NFB and other transcription factor alterations in relation to internucleosomal DNA fragmentation after the intrastriatal administration of the potent NMDA receptor agonist QA. We also studied
the effect of inhibiting NFB activity on QA-induced apoptosis using
a cell-permeable recombinant peptide (NFB SN50) to block NFB
nuclear translocation. The results indicate that a complex pattern of
transcription factor change precedes the appearance of internucleosomal
DNA fragmentation and most noticeably that the activation of NFB may
contribute to the QA-induced apoptosis of striatal neurons.
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Materials and Methods |
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Summary
Introduction
Materials & Methods
Results
Discussion
References
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Animals. Sprague-Dawley rats weighing 300-350 g were purchased from Taconic. They were housed two per cage in a standard animal room with a 12-hr light/dark cycle and given free access to food and water. All procedures were conducted in accordance with National Institutes of Health Guidelines for the Care and Use of Laboratory Animals.
Drug treatment. To study the time course of QA-induced DNA fragmentation, rats were unilaterally infused with QA (120 nmol) or saline into the striatum (7) and killed 2, 6, 12, or 24 hr later. Whole brains were used for tissue sections for TUNEL assay or striata were dissected for the extraction of genomic DNA.
To examine the time course of QA-induced changes in transcription factor binding activity, rats were infused with QA (120 nmol) and killed 2, 6, 12, or 24 hr later. Their striata were dissected on a cold plate for nuclear protein extraction. To evaluate the effect of NMDA receptor blockade on QA-induced alterations in transcription factors, animals were given MK-801 (dizocilpine; RBI, Natick, MA) intraperitoneally. The first MK-801 dose (2.0 mg/kg) was injected 15 min before and the second (2.0 mg/kg) and third doses (1.0 mg/kg) 3 and 6 hr after intrastriatal QA infusion. Control rats received either intraperitoneal injections of vehicle (0.9% NaCl) plus intrastriatal QA or intraperitoneal injections of MK-801 plus intrastriatal vehicle (0.9% NaCl). Animals were killed 12 hr after striatal QA or vehicle infusion, and their striata were dissected for nuclear protein extraction. To study the effect of a protein synthesis inhibitor on QA-induced transcription factor alterations, rats were injected intrastriatally with CHX (480 nmol in 2 µl of 0.9% NaCl; Sigma, St. Louis, MO). Fifteen minutes later, QA was injected intrastriatally as previously described. Control animals received either intrastriatal vehicle plus intrastriatal QA or intrastriatal CHX plus intrastriatal vehicle. Animals were killed 12 hr after striatal QA or vehicle administration, and their striata were dissected for nuclear protein extraction. To assess the effect of a cell permeable recombinant peptide that inhibits NFB nuclear translocation (NFB SN50; Biomol, Plymouth
Meeting, PA) on QA-induced NFB activation and internucleosomal DNA
fragmentation, rats received either a single intrastriatal injection of
NFB SN50 (20 µg) 15 min before QA treatment, or two injections of
NFB SN50 (10 or 30 µg/injection) 15 min before and 8 hr after QA
treatment. Animals were killed 12 or 24 hr after QA administration, and
their striata were dissected for nuclear protein and genomic DNA
extraction. The effect of NFB SN50 on QA-induced striatal cell death
was examined in animals given a single injection of NFB SN50 (20 µg) 15 min before QA treatment, or two injections of NFB SN50 (10 or 30 µg/injection) 15 min before and 8 hr after QA treatment.
Animals were killed 10 days later, and brain sections were processed
for receptor autoradiography and in situ hybridization
histochemistry.
Genomic DNA isolation and electrophoresis.
Genomic DNA was
isolated (Qin et al., 1996), and 20 µg were
electrophoresed on 2% agarose gel (NuSieve 3:1) for 3 hr. DNA fragments were detected with a UV transilluminator after staining with
ethidium bromide.
TUNEL. Brain sections (12 mm) were cut on a cryostat and thaw-mounted onto gelatin-coated microslides. Sections were fixed in phosphate-buffered 10% formalin for 10 min and then rinsed three times in phosphate-buffered saline, pH 7.4. DNA fragmentation was evaluated in individual cells using an apoptosis detection kit (ApopTag; Oncor, Gaithersburg, MD) according to the manufacturer's protocol. DNA fragmentation was disclosed by fluorescence microscopy and photographed.
Nuclear protein preparation and gel shift assay.
Nuclear
proteins were extracted by a modification of a previously described
procedure (Ogita and Yoneda, 1994) and dialyzed using microdialyzers
(Daigger, Wheeling, IL). Double-stranded oligodeoxynucleotide
containing consensus sequences for different transcription factors were
purchased from Santa Cruz Biotechnology (Santa Cruz, CA) and Promega
(Madison, WI). Double-stranded DNA probes were labeled with
[32P]ATP by T4 polynucleotide kinase (Promega).
Nuclear protein (5-14 µg) were incubated with radioactively labeled
DNA probes (about 40,000 cpm) for 15 min at room temperature in a
binding buffer containing 5 mM MgCl2,
2.5 mM EDTA, 25 mM dithiothreitol, 250 mM NaCl, 50 mM Tris·HCl, pH 7.5, 0.25 mg/ml
poly(dI·dC) and 20% glycerol (Promega). Nuclear proteins were mixed
with 1/10 volume of a loading buffer containing 250 mM
Tris·HCl (pH 7.5), 0.2% bromphenol blue, 0.2% xylene cyanol, and
40% glycerol and then electrophoresed on 4% polyacrylamide gel
(monoacrylamide/bisacrylamide, 80:1) with 0.5 × Tris/borate/EDTA
(1× = 89 mM Tris base, 89 mM boric acid, and 2 mM Na2-EDTA). Autoradiograms were developed by
exposing vacuum-dried gels to x-ray film at 
80° with intensifying screens for 12-48 hr. The specificity of transcription factor binding
to DNA probes used in the gel shift assay was tested by assessing the
affinity of the nuclear proteins to individual
32P-labeled transcription factor consensus
sequences in the presence of an excess of the unlabeled probes, or by
mutation of DNA-protein binding motifs in DNA probes. The specificity
of NFB binding was also confirmed by supershift assay using p65
(NFBp65, sc-109; Santa Cruz Biotechnology) and p50 (NFBp50,
sc-1190, Santa Cruz Biotechnology) antibodies. Autoradiographic results
were semiquantitatively evaluated by means of an image analyzer (Image
1.42; National Institutes of Health). A standard template was used to
ensure that the binding signal from each sample was measured in the
same sized area. Nonspecific binding was determined by adding a 60-fold excess of unlabeled DNA probes to the assay. Specific binding was
calculated by subtracting nonspecific binding from total binding.
Receptor autoradiography and in situ hybridization
histochemistry.
Receptor autoradiography was performed as
previously described (Qin et al., 1994a). Briefly, brain
sections were incubated in 50 mM Tris·HCl buffer, pH 7.4, containing 2 nM [3H]SCH-23390 (NEN,
Boston, MA) and 80 nM ketanserin for 1 hr at room
temperature. Nonspecific binding was determined by incubating adjacent
sections in the buffer with 2 µM SCH-23390 added.
Sections were rinsed, air-dried, and exposed to x-ray film (Hyperfilm, [3H] Sensitive, Amersham) for 10 days. The
density of D1 dopamine receptors was determined
using an image analyzer. Three brain sections (bregma 1.7 to 0.3)
from each treated animal were analyzed. In situ
hybridization histochemistry was performed as described previously with
minor modifications (Qin et al., 1994b). A 36-mer oligonucleotide probe (5
-GCT AAA CCA ATG ATA TCC AAA CCA GTA GAG AGC
TGG-3) complimentary to rat GAD mRNA encoding a 67-kDa GAD protein was
synthesized by Genosys. Oligonucleotide probes were labeled with
[33P]dATP using terminal deoxynucleotidyl
transferase and purified by filtration chromatography (Chroma Spin-10,
Clontech). Formalin-fixed sections were incubated with labeled probes
in a hybridization cocktail (Amresco, Solon, OH) at 37° for 18 hr.
After hybridization, sections were washed, dehydrated , and exposed to
x-ray film (Hyperfilm Max; Amersham) for 10 days. The results were
quantitatively assessed with an image analyzer. Three brain sections
(bregma 1.7 to 0.3) from each treated animal were analyzed.
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Results |
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Top
Summary
Introduction
Materials & Methods
Results
Discussion
References
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Temporal pattern of QA-induced DNA fragmentation. DNA fragmentation first became detectable on ethidium bromide-stained agarose gels 12 hr after QA administration. DNA fragmentation was further increased 24 hr after QA treatment. The size of the DNA fragments approximated multimers of 180-200 base pairs and thus formed typical DNA ladders (Fig. 1). Internucleosomal DNA fragmentation was not observed in the contralateral striatum or in the vehicle injected striatum (data not shown).
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Temporal pattern of QA-induced changes in striatal transcription
factor binding.
The intrastriatal administration of QA
significantly altered binding activities of four of the seven
transcription factors studied (Figs. 3
and 4). QA induced a rapid rise in AP-1
binding, which peaked 6 hr after excitotoxin injection and then slowly returned to basal levels. NFB binding, which remained low in the
vehicle-treated striatum, increased markedly in the QA-treated striatum. The maximal increase occurred 12 hr after QA infusion. Subsequently, NFB binding gradually diminished, although continuing to be significantly elevated 24 hr after QA treatment. A small transient rise in E2F-1 binding activity was observed 6 hr after the QA
treatment. In contrast, QA induced a gradual decline in OCT-1 binding,
with the decrement reaching statistical significance 12 hr after
excitotoxin exposure. SP-1 binding activity also decreased, although
only briefly 12 hr after QA administration. There was no appreciable
change in CREB or Myc-Max binding activity.
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NMDA receptor antagonist and protein synthesis inhibitor effects on
QA-induced alterations in transcription factor binding.
Given
alone, systemically administered MK-801 significantly inhibited SP-1
binding activity (p < 0.001), but had no
effect on the other striatal transcription factors studied. MK-801
co-administration completely blocked the QA-induced increases in AP-1
(p < 0.05) and NFB
(p < 0.05) binding found 12 hr after
intrastriatal excitotoxin infusion. MK-801 also tended to reverse the
QA-induced decrease in OCT-1 binding. Although MK-801 had an inhibitory
effect on SP-1, there was no additive effect on the QA-induced decline
in SP-1 binding activity when it was co-administered with QA (Fig. 5).
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B binding (p < 0.01) slightly,
although significantly, whereas it reduced SP-1 binding activity. CHX
co-administration attenuated rather than potentiated the QA-induced
increases in NFB binding (p < 0.05). A
tendency for CHX to diminish the QA-induced rise in AP-1 binding and
reverse the decrement in OCT-1 binding activity did not quite attain
statistical significance. CHX had no effect on the QA-induced reduction
in SP-1 binding activity (Fig. 6).
|
NFB antagonist effects on QA-induced internucleosomal DNA
fragmentation and striatal cell death.
Co-administration of the
NFB inhibitor, NFB SN50, markedly reduced the QA-induced
activation of NFB (p <0.001), but did not alter
QA-induced increases in AP-1 binding. NFB SN50 had no significant
effect on QA-induced reductions in OCT-1 and SP-1 binding (Fig.
7). Treatment with two doses of NFB
SN50 (10 or 30 µg/dose) attenuated QA-induced internucleosomal DNA
fragmentation 24 hr after QA treatment in a dose-dependent manner.
NFB SN50 alone did not produce appreciable internucleosomal DNA
fragmentation (Fig. 8A). A single dose of
NFB SN50 (20 µg) also substantially inhibited QA-induced
internucleosomal DNA fragmentation (Fig. 8B). Similarly, a single
injection of NFB SN50 (20 µg) significantly attenuated the
QA-induced decrease in striatal D1 dopamine
receptor binding sites (p < 0.001, Fig.
9A). In situ hybridization
histochemistry further confirmed that a single dose of NFB SN50 (20 µg) reduced the QA-induced decrement in striatal
GAD67 mRNA levels (p < 0.001, Fig. 9B). However, a second injection (20 µg) of NFB SN50
failed to provide additional protection against QA-induced striatal
cell death as indicated both by receptor autoradiography and in
situ hybridization histochemistry.
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Discussion |
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Top
Summary
Introduction
Materials & Methods
Results
Discussion
References
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The present results document the complex temporal relation between
acute excitotoxin exposure and transcription factor induction in rat
striatum. After QA administration, AP-1 and NFB rapidly increased,
whereas OCT-1 and SP-1 gradually declined. These transcription factor
alterations, detected by a highly sequence-specific mobility shift
assay, are consistent with previously reported changes in immediate
early gene expression (including c-fos, fos B,
c-jun, jun B) as well as alterations in AP-1 and
NFB binding activities in neurons as a consequence of glutamate
receptor activation (Coyle et al., 1989; Bading et
al., 1993; Dure et al., 1995; Guerrini et
al., 1995; Kaltschmidt et al., 1995).
The excitotoxin-induced effects on striatal transcription factors
observed in this study were probably mediated by glutamate receptors of
the NMDA subtype, because QA, at the doses used, acts selectively at
these receptors. Moreover, the noncompetitive NMDA receptor antagonist
MK-801, in amounts previously found to block QA-induced cell death by a
process having the hallmarks of apoptosis, totally prevented QA
associated changes in AP-1, NFB, and OCT-1 binding. Our results with
MK-801, which inhibits calcium permeability when bound to NMDA
channels, are thus consistent with earlier reports suggesting that
enhanced calcium influx, as a consequence of NMDA channel activation,
contributes to the observed alterations in transcription factor binding
(Dure et al., 1995).
The observed QA-induced increases in NFB and AP-1 were attenuated by
the protein synthesis inhibitor CHX. Previously, we have reported that
CHX, under the conditions used in this study, diminishes QA-induced
internucleosomal DNA fragmentation (Qin et al., 1996). Thus
although not all types of apoptosis depend on new protein synthesis
(Milligan et al., 1994), the present results suggest a
genetic program, involving certain transcription factor inductions, at
least in part, attends the excitotoxic death of striatal neurons.
However, CHX only modestly, although significantly, attenuated
QA-induced internucleosomal DNA fragmentation and NFB activation in
our studies. This may reflect the fact that a single dose of CHX does
not completely inhibit new protein synthesis, or that newly synthesized
proteins contribute little to the apoptotic process. In the case of the
NFB family, presynthesized proteins in association with an
inhibitory protein IB normally reside in the cytoplasm. Upon
appropriate stimulation, IB is degraded and NFB can then
translocate to the nucleus and regulate gene expression.
Transcription factor changes found in this study presumably relate to
the excitotoxic induction of an apoptotic cascade in striatal neurons.
Although a significant reactive glia contribution to these
transcription factor alterations cannot be excluded, previous
investigations have suggested that the QA-induced death of medium spiny
neurons, which account for more than 90% of nerve cells in rat
striatum, involves an NMDA receptor-mediated apoptotic mechanism. For
example, QA has not only been observed to produce many of the hallmarks
of apoptosis, including internucleosomal DNA fragmentation, chromatin
condensation, and nuclear fragmentation (Qin et al., 1996),
but also to induce proteins, such as p53 and Bax, known to be directly
involved in the apoptotic process (Hughes et al., 1996).
Using the same QA administration technique, we now find a marked
increase in the number of TUNEL-positive nuclei and clear laddering of
DNA fragments on agarose gels 12-24 hr after excitotoxin exposure.
These fragments were of a size, multimers of 180-200 base pairs,
expected from DNA cleavage by endonuclease during apoptosis. Moreover,
all apoptotic stigmata appeared in close temporal relation to the
observed transcription factor alterations: maximal induction of AP-1
and NFB binding activity occurred 6-12 hr after QA treatment, thus
preceding internucleosomal DNA fragmentation, whereas OCT-1 binding had
significantly declined when DNA fragmentation peaked. In addition,
MK-801 and CHX doses that inhibited QA-induced changes in transcription
factors were the same as those that reduced apoptosis in our earlier
study (Qin et al., 1996).
The present results further suggest that NFB plays an important role
in QA-induced apoptosis. The functional consequences of preventing
NFB activation were evaluated by means of NFB SN50, which
interferes with NFB nuclear translocation. This recombinant peptide
contains the nuclear localization signal of the transcription factor
NFB p50 and the hydrophobic region of Kaposi fibroblast growth
factor. The Kaposi fibroblast growth factor hydrophobic region confers
cell permeability, whereas the nuclear localization signal inhibits
translocation of the NFB complex from the cytoplasm to the nucleus.
Lin et al. (1995b) have demonstrated that this peptide
inhibits nuclear translocation of NFB in vitro in a
dose-dependent manner. In the present studies, co-administration of the
recombinant cell-permeable peptide selectively inhibited both
QA-induced NFB activation and internucleosomal DNA fragmentation.
Moreover, the decrement in internucleosomal DNA fragmentation was
associated with a reduction in striatal cell death, as indicated by
both receptor autoradiography and in situ hybridization
histochemistry. It should be noted, however, that inhibition of NFB
activation failed to protect all striatal neurons against QA toxicity.
Conceivably, additional NFB-independent apoptotic cascades exist or
both apoptosis and necrosis contribute to excitotoxic neuronal
destruction. Alternatively, a basal level of NFB activity may be
required for the maintenance of normal cell function, although support
for this possibility is somewhat weakened by the fact that NFB SN50
alone can cause tissue damage.
The participation of transcription factors in neuronal apoptosis has
been studied in various experimental systems. Proteins that bind to
AP-1 consensus sequences include the Fos and Jun families; increases in
both c-jun and c-fos proteins or mRNAs have been described during the
apoptotic death of neurons (Dure et al., 1995; Estus
et al., 1994; Tong and Perez-Polo, 1995). Similarly,
intrastriatal injections of QA increase the expression of c-fos mRNA in
areas which degenerate, and especially in medium spiny neurons (Aronin
et al., 1991). A study showing that anti-c-Jun antibodies
block apoptosis in cultured sympathetic neurons provides relatively
direct evidence of transcription factor involvement in the apoptotic
process (Estus et al., 1994). Binding activity of the OCT-1
transcription factor family, implicated in the regulation of various
housekeeping genes and certain cell-specific genes, reportedly also
decline in ischemia-induced brain damage and nerve growth factor
deprivation-induced apoptosis in cultured cells (Wang and Pittman,
1993).
NFB is well known to be involved in regulating important cellular
functions including programmed cell death. Although somewhat inconsistent results have been reported, NFB activation generally inhibits apoptosis, particularly when induced by tumor necrosis factor
(Beg et al., 1993; Wu and Lozano, 1994). Although convincing evidence in support of the participation of NFB in apoptotic mechanisms within neuronal tissues has not been previously reported, NFB has been implicated in the programmed death of cultured cells (Grimm et al., 1996; Lin et al., 1995a). The
present study provides the first in vivo results indicating
that NFB may contribute to excitotoxin-induced apoptosis of striatal
medium spiny neurons. Conceivably, NFB influences apoptotic
mechanisms in neurons differently than in other cells. Whether the
apparently pro-apoptotic role of NFB we observed here in medium
spiny neurons can be generalized to other types of neurons or to other
apoptotic triggers remains to be determined. Nevertheless, the results
of this study, taken together with recent findings suggesting that
aspirin and sodium salicylate protect cultured cerebellar granule cells
and hippocampal neurons against glutamate toxicity by a mechanism
possibly involving NFB inhibition (Grilli et al., 1996),
could have important implications for the treatment of striatal
neurodegenerative disorders.
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Footnotes |
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Received July 21, 1997; Accepted September 13, 1997
Send reprint requests to: Thomas N. Chase, M.D., Chief, ETB NINDS, NIH, Bldg. 10, Room 5C103, 10 Center Drive, MSC 1406, Bethesda, MD 20892-1406. E-mail: chase{at}helix.nih.gov
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Abbreviations |
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HD, Huntington's disease;
QA, quinolinic
acid;
CHX, cycloheximide;
TUNEL, terminal transferase-mediated
dUTP-digoxigenin nick end labeling;
NMDA, N-methyl-D-aspartate;
ANOVA, analysis of
variance;
IB, inhibitor B;
NFB, nuclear factor-B;
AP-1, activator protein 1;
CREB, cAMP response element binding protein.
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References |
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Top
Summary
Introduction
Materials & Methods
Results
Discussion
References
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 B. R. Ryu, Y. A. Lee, S. J. Won, J.-H. Noh, S.-Y. Chang, J.-M. Chung, J. S. Choi, C. K. Joo, S. H. Yoon, and B. J. Gwag The Novel Neuroprotective Action of Sulfasalazine through Blockade of NMDA Receptors J. Pharmacol. Exp. Ther., April 1, 2003; 305(1): 48 - 56. [Abstract] [Full Text]
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 V. Poulaki, C. S. Mitsiades, A. M. Joussen, A. Lappas, B. Kirchhof, and N. Mitsiades Constitutive Nuclear Factor-{kappa}B Activity Is Crucial for Human Retinoblastoma Cell Viability Am. J. Pathol., December 1, 2002; 161(6): 2229 - 2240. [Abstract] [Full Text] [PDF]
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 N. Mitsiades, C. S. Mitsiades, V. Poulaki, D. Chauhan, P. G. Richardson, T. Hideshima, N. Munshi, S. P. Treon, and K. C. Anderson Biologic sequelae of nuclear factor-kappa B blockade in multiple myeloma: therapeutic applications Blood, May 13, 2002; 99(11): 4079 - 4086. [Abstract] [Full Text] [PDF]
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 L. Acarin, B. Gonzalez, and B. Castellano Triflusal Posttreatment Inhibits Glial Nuclear Factor-{kappa}B, Downregulates the Glial Response, and Is Neuroprotective in an Excitotoxic Injury Model in Postnatal Brain Stroke, October 1, 2001; 32(10): 2394 - 2402. [Abstract] [Full Text] [PDF]
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 M. Aoki, T. Nata, R. Morishita, H. Matsushita, H. Nakagami, K. Yamamoto, K. Yamazaki, M. Nakabayashi, T. Ogihara, and Y. Kaneda Endothelial Apoptosis Induced by Oxidative Stress Through Activation of NF-{kappa}B: Antiapoptotic Effect of Antioxidant Agents on Endothelial Cells Hypertension, July 1, 2001; 38(1): 48 - 55. [Abstract] [Full Text] [PDF]
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 L. Li, J. N. Rao, B. L. Bass, and J.-Y. Wang NF-{kappa}B activation and susceptibility to apoptosis after polyamine depletion in intestinal epithelial cells Am J Physiol Gastrointest Liver Physiol, May 1, 2001; 280(5): G992 - G1004. [Abstract] [Full Text] [PDF]
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Z.-H. Qin, Y. Wang, R.-W. Chen, X. Wang, M. Ren, D.-M. Chuang, and T. N. Chase Prostaglandin A1 Protects Striatal Neurons against Excitotoxic Injury in Rat Striatum J. Pharmacol. Exp. Ther., April 1, 2001; 297(1): 78 - 87. [Abstract] [Full Text]
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 C. W. Xiao, K. Ash, and B. K. Tsang Nuclear Factor-{{kappa}}B-Mediated X-Linked Inhibitor of Apoptosis Protein Expression Prevents Rat Granulosa Cells from Tumor Necrosis Factor {{alpha}}-Induced Apoptosis Endocrinology, February 1, 2001; 142(2): 557 - 563. [Abstract] [Full Text] [PDF]
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 Y. Huang, K. R. Johnson, J. S. Norris, and W. Fan Nuclear Factor-{{kappa}}B/I{{kappa}}B Signaling Pathway May Contribute to the Mediation of Paclitaxel-induced Apoptosis in Solid Tumor Cells Cancer Res., August 1, 2000; 60(16): 4426 - 4432. [Abstract] [Full Text]
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 H. Matsushita, R. Morishita, T. Nata, M. Aoki, H. Nakagami, Y. Taniyama, K. Yamamoto, J. Higaki, K. Yasufumi, and T. Ogihara Hypoxia-Induced Endothelial Apoptosis Through Nuclear Factor-{kappa}B (NF-{kappa}B)-Mediated bcl-2 Suppression : In Vivo Evidence of the Importance of NF-{kappa}B in Endothelial Cell Regulation Circ. Res., May 12, 2000; 86(9): 974 - 981. [Abstract] [Full Text] [PDF]
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S. L. Hickenbottom, J. C. Grotta, R. Strong, L. A. Denner, J. Aronowski, and R. L. Macdonald Nuclear Factor-{kappa}B and Cell Death After Experimental Intracerebral Hemorrhage in Rats • Editorial Comment Stroke, November 1, 1999; 30 (11): 2472 - 2478. [Abstract] [Full Text] [PDF]
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 Y. Luo, A. Hattori, J. Munoz, Z.-H. Qin, and G. S. Roth Intrastriatal Dopamine Injection Induces Apoptosis Through Oxidation-Involved Activation of Transcription Factors AP-1 and NF-kappa B in Rats Mol. Pharmacol., August 1, 1999; 56(2): 254 - 264. [Abstract] [Full Text]
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 S. I. Savitz and D. M. Rosenbaum Review : Gene Expression after Cerebral Ischemia Neuroscientist, July 1, 1999; 5(4): 238 - 253. [Abstract] [PDF]
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 Z.-H. Qin, R.-W. Chen, Y. Wang, M. Nakai, D.-M. Chuang, and T. N. Chase Nuclear Factor kappa B Nuclear Translocation Upregulates c-Myc and p53 Expression during NMDA Receptor-Mediated Apoptosis in Rat Striatum J. Neurosci., May 15, 1999; 19(10): 4023 - 4033. [Abstract] [Full Text] [PDF]
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R. Y. Liu, C. Fan, N. E. Olashaw, X. Wang, and K. S. Zuckerman Tumor Necrosis Factor-alpha -induced Proliferation of Human Mo7e Leukemic Cells Occurs via Activation of Nuclear Factor kappa B Transcription Factor J. Biol. Chem., May 14, 1999; 274(20): 13877 - 13885. [Abstract] [Full Text] [PDF]
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J. W. Choi and S. E. Jung Lovastatin-Induced Proliferation Inhibition and Apoptosis in C6 Glial Cells J. Pharmacol. Exp. Ther., April 1, 1999; 289(1): 572 - 579. [Abstract] [Full Text]
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 T. R. Torgerson, A. D. Colosia, J. P. Donahue, Y.-Z. Lin, and J. Hawiger Regulation of NF-{kappa}B, AP-1, NFAT, and STAT1 Nuclear Import in T Lymphocytes by Noninvasive Delivery of Peptide Carrying the Nuclear Localization Sequence of NF-{kappa}B p50 J. Immunol., December 1, 1998; 161(11): 6084 - 6092. [Abstract] [Full Text] [PDF]
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R. Y. Liu, C. Fan, G. Liu, N. E. Olashaw, and K. S. Zuckerman Activation of p38 Mitogen-activated Protein Kinase Is Required for Tumor Necrosis Factor-alpha -supported Proliferation of Leukemia and Lymphoma Cell Lines J. Biol. Chem., July 7, 2000; 275(28): 21086 - 21093. [Abstract] [Full Text] [PDF]
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K. Pahan, F. G. Sheikh, X. Liu, S. Hilger, M. McKinney, and T. M. Petro Induction of Nitric-oxide Synthase and Activation of NF-kappa B by Interleukin-12 p40 in Microglial Cells J. Biol. Chem., March 9, 2001; 276(11): 7899 - 7905. [Abstract] [Full Text] [PDF]
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