Previous Article | Next Article 
Volume 17, Number 11,
Issue of June 1, 1997
pp. 4212-4222
Copyright ©1997 Society for Neuroscience
Alzheimer's Presenilin Mutation Sensitizes Neural Cells to
Apoptosis Induced by Trophic Factor Withdrawal and Amyloid
-Peptide: Involvement of Calcium and Oxyradicals
Qing Guo1,
Bryce L. Sopher2,
Katsutoshi Furukawa1,
Dao G. Pham2,
Nic Robinson1,
George M. Martin2, and
Mark P. Mattson1
1 Sanders-Brown Research Center on Aging and Department
of Anatomy and Neurobiology, University of Kentucky, Lexington,
Kentucky 40536, and 2 Department of Pathology, University
of Washington, Seattle, Washington 98195-7470
ABSTRACT
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- DISCUSSION
- FOOTNOTES
- REFERENCES
ABSTRACT 
Most autosomal dominant inherited forms of early onset Alzheimer's
disease (AD) are caused by mutations in the presenilin-1 (PS-1) gene on
chromosome 14. PS-1 is an integral membrane protein with six to nine
membrane-spanning domains and is expressed in neurons throughout the
brain wherein it is localized mainly in endoplasmic reticulum (ER). The
mechanism or mechanisms whereby PS-1 mutations promote neuron
degeneration in AD are unknown. Recent findings suggest links among
deposition of amyloid 
-peptide (A), oxidative stress, disruption
of ion homeostasis, and an apoptotic form of neuron death in AD. We now
report that expression of the human PS-1 L286V mutation in PC12 cells
increases their susceptibility to apoptosis induced by trophic factor
withdrawal and A. Increases in oxidative stress and intracellular
calcium levels induced by the apoptotic stimuli were exacerbated
greatly in cells expressing the PS-1 mutation, as compared with control cell lines and lines overexpressing wild-type PS-1. The antiapoptotic gene product Bcl-2 prevented apoptosis after NGF withdrawal from differentiated PC12 cells expressing mutant PS-1. Elevations of [Ca2+]i in response to thapsigargin, an
inhibitor of the ER Ca2+-ATPase, were increased in cells
expressing mutant PS-1, and this adverse effect was abolished in cells
expressing Bcl-2. Antioxidants and blockers of calcium influx and
release from ER protected cells against the adverse consequences of the
PS-1 mutation. By perturbing cellular calcium regulation and promoting
oxidative stress, PS-1 mutations may sensitize neurons to apoptotic
death in AD.
Key words:
Alzheimer's disease;
antioxidant;
bcl-2;
dantrolene;
endoplasmic reticulum;
fura-2;
nerve growth factor
INTRODUCTION
Alzheimer's disease (AD) is a progressive and
always fatal neurodegenerative disorder characterized by the death of
neurons in brain regions involved in learning and memory processes (for review, see Selkoe, 1989
). Accumulations of insoluble fibrillar aggregates of a protein called amyloid -peptide (A) are
implicated in the pathogenesis of AD. A is associated with
degenerating neurons in AD brain, and mutations in the gene for the
amyloid precursor protein (the source of A) cause a small percentage of cases of inherited familial AD (for review, see Mullan and Crawford,
1993). Moreover, mice genetically engineered to produce mutated human
amyloid precursor protein exhibit A deposition and cognitive
impairments (Games et al., 1995; Hsaio et al., 1996). A damages and
kills cultured neurons by a mechanism involving oxidative stress and
disruption of cellular calcium homeostasis (Mattson et al., 1992; Behl
et al., 1994; Fukuyama et al., 1994; Goodman and Mattson, 1994; Mark et
al., 1995, 1997a). Apoptosis is a form of "programmed" cell death
characterized by plasma membrane phospholipid alterations, cell
shrinkage, and nuclear DNA condensation and fragmentation (Bredesen,
1995; Thompson, 1995). A induces apoptosis in cultured neurons
(Forloni et al., 1993; Loo et al., 1993), and studies of postmortem
brain tissue suggest that neuronal apoptosis occurs in AD (Su et al.,
1994; Smale et al., 1995). The molecular and cellular mechanisms that
predispose neurons to apoptotic death in AD are unknown.
An important step toward elucidating the cause or causes of neuron
degeneration in AD was made with the identification of the genes
responsible for the majority of cases of autosomal dominant inherited
forms of early onset AD. The genes, called presenilin-1 (PS-1;
chromosome 14) and presenilin-2 (PS-2; chromosome 1), encode proteins
predicted to be integral membrane proteins with six to nine
membrane-spanning domains (Levy-Lahad et al., 1995; Rogaev et al.,
1995; Sherrington et al., 1995; Doan et al., 1996; Li and Greenwald,
1996). Immunohistochemical analyses indicate that PS-1 is expressed in
neurons throughout the brain (Cook et al., 1996; Cribbs et al., 1996;
Elder et al., 1996); PS-1 has been localized to both degenerating
(Murphy et al., 1996) and nondegenerating (Giannakopoulos et al., 1997)
neurons in AD brain. In cultured cells PS-1 localizes to subcellular
compartments and seems to be at particularly high levels in the
endoplasmic reticulum (ER) (Kovacs et al., 1996; Walter et al., 1996).
PS-1 can be processed endoproteolytically (Thinakaran et al., 1996),
although the significance of such processing is unknown. Because PS-1
mutations account for the majority of cases of inherited early onset
forms of AD, understanding the normal functions of PS-1 and how PS-1
mutations promote neuron degeneration are central issues in the AD
field. We report that expression of a human PS-1 mutation in PC12 cells increases their vulnerability to apoptosis induced by trophic factor
withdrawal and A. The mechanism whereby mutant PS-1 promotes apoptosis seems to involve disruption of calcium homeostasis and increased oxidative stress.
MATERIALS AND METHODS
Expression of wild-type and mutant PS-1 in PC12 cells.
Rat pheochromocytoma (PC12) cells (Black and Greene, 1982) were
maintained at 37°C (5% CO2 atmosphere) in RPMI-1640
medium supplemented 10% with heat-inactivated horse serum and 5% with
heat-inactivated fetal bovine serum. A full-length human PS-1 cDNA and
a PS-1 cDNA containing the L286V mutation were cloned into either the
expression vector pTRE in the Tet-off expression system (Clontech,
Cambridge, UK) or pRc/CMV to produce pTRE-PS1 and pTRE-PS1L286V or
pCMV-PS1 and pCMV-PS1L286V, where, in both cases, the expression of
PS-1 and PS-1 L286V cDNAs is under the control of CMV promoter. PC12 cells were transfected with Lipofectamine (Life Technologies, Gaithersburg, MD). Stable expression of PS-1 L286V in PC12 cells with
the pRc/CMV vector did not affect cell viability significantly during
the G418 selection procedure. Double-stable PC12 cell lines in which
PS-1 and PS-1 L286V expression could be suppressed or induced were
established by first generating stable lines expressing the Tet-off
system (cells were selected for 4 weeks in the presence of 0.8 mg/ml of G418). G418-resistant clones were isolated and cotransfected with the response plasmids pTRE-PS1 or pTRE-PS1L286V and
pTK-Hyg. Cells were grown in selection medium containing 0.4 mg/ml hygromycin, and stable clones were isolated after 4 weeks and screened for PS-1 expression in the presence or absence of 2 µg/ml Tet by Western blot or RT-PCR. Resulting double-stable cell lines that exhibited high levels of expression after removal of
tetracycline were used for subsequent experiments. A PC12 cell line
overexpressing Bcl-2 (a generous gift from D. Bredesen, The Burnaham
Institute, La Jolla, CA; Kane et al., 1993) was used to generate stable
lines expressing the Tet-off system as described above. The latter
lines then were cotransfected with the response plasmids pTRE-PS1 or
pTRE-PS1L286V and pTK-Hyg, and stable lines exhibiting high levels of
expression of wild-type and mutant PS-1 were used for experiments.
RT-PCR analysis was performed as described previously (Guo et al.,
1996). Briefly, mRNA from the cultured cells was isolated and
reverse-transcribed via the reverse transcription system (Promega, Madison, WI) and the 3
primer 5-GCTTCCCATTCCTCACTGAA-3. cDNA (2.5 µl) was used as a template in a 50 µl PCR, using 15-40 cycles of
94°C (1 min), 60°C (2 min), and 72°C (2 min) with a final
extension time of 10 min at 72°C. Reaction mixtures were as
recommended for Taq polymerase (Perkin-Elmer Cetus, Oak
Brook, IL), except that Taq was added after the mixtures
were heated to 95°C for 7 min. The 3 primer used was the
oligonucleotide used to prime the cDNA synthesis; the 5 primer was 5-
GTGGCTGTTTTGTGTCCGAA-3. The PCR products were resolved and visualized
by electrophoresis in 3% agarose gel stained with ethidium bromide.
Because the Leu to Val mutation at codon 286 creates a PvuII
site, the wild-type RT-PCR product could not be cut by PvuII
and generated a single 251-bp fragment, whereas the mutation resulted
in PvuII cleavage of the product into 79 and 172 bp
fragments.
Experimental treatments. Cells were differentiated to a
neuron-like phenotype by incubation in medium with reduced serum
concentration (2% fetal bovine serum) and containing 50 ng/ml
nerve growth factor (NGF) (Black et al., 1982). Immediately before
experimental treatment the medium was replaced with Locke's solution
containing (in mM): NaCl 154, KCl 5.6, CaCl2
2.3, MgCl2 1.0, NaHCO3 3.6, glucose 5, and
HEPES 5, pH 7.2. Serum withdrawal from undifferentiated PC12 cells and
NGF withdrawal from differentiated PC12 cells were accomplished by
repeated washing of cells with Locke's solution. Synthetic A25-35
was purchased from Bachem (Torrence, CA), and stocks were prepared at a
concentration of 1 mM in water and allowed to incubate overnight at 37°C before addition to cultures. Nifedipine, sodium dantrolene, thapsigargin, vitamin E, and propyl gallate were purchased from Sigma (St. Louis, MO) and prepared as 500× stocks in ethanol.
Generation of PS-1 antibodies and Western blot analysis.
Affinity-purified polyclonal antibody was isolated from serum of rabbits injected with a synthetic peptide with a sequence
(NH2-NDDGGFSEEWEAQRD-COOH) corresponding to amino acids
331-345 of the loop region of human PS-1. Preliminary studies showed
that this antibody recognizes both wild-type and PS-1 L286V. For
Western blot analysis solubilized cell proteins were separated by
electrophoresis in a 12% polyacrylamide gel, transferred to a
nitrocellulose sheet, and immunoreacted with PS-1 antibody (1:100). The
nitrocellulose sheet was processed further with HRP-conjugated
anti-mouse secondary antibody and a chemiluminescence detection method
(Amersham, Arlington Heights, IL).
Analyses of cell death and apoptosis. Quantification of LDH
levels in culture medium was done as described previously (Bruce et
al., 1996). Levels of cellular
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)
reduction, a measure of mitochondrial redox status and function
(Shearman et al., 1994), were quantified as described previously
(Mattson et al., 1995). Briefly, MTT solution (5 mg/ml PBS) was
added to cultures (1:10, v:v; MTT solution/culture medium) and allowed
to incubate for 3 hr. The cells were washed three times with Locke's
solution and solubilized in dimethylsulfoxide; the absorbance in each
culture well was quantified with a plate reader. Methods used to
establish apoptotic cell death included Hoescht and propidium iodide
staining of DNA and suppression of cell death by macromolecular
synthesis inhibitors. For Hoescht and propidium iodide staining, cells
were fixed in 4% paraformaldehyde, membranes were permeabilized with
0.2% Triton X-100, and cells were stained with the fluorescent
DNA-binding dyes Hoescht 33342 or propidium iodide, as described
previously (Mark et al., 1995; Kruman et al., 1996). Hoescht-stained
cultures were used to quantify the percentage of "apoptotic" cells;
cells with condensed and fragmented DNA were considered apoptotic,
whereas cells in which the DNA was distributed diffusely and uniformly
throughout the nucleus were considered not apoptotic. Cells were
visualized under epifluorescence illumination (340 nm excitation and
510 nm barrier filter) with a 40× oil immersion objective. Cells were
counted in four random 40× fields per culture; counts were made
without knowledge of cell type or treatment history. Images of
propidium iodide-stained cells were acquired with a confocal laser
scanning microscope (488 nm excitation and 510 nm barrier filter;
Molecular Dynamics, Sunnyvale, CA) with a 60× oil immersion
objective.
Measurements of intracellular peroxide and calcium levels.
Peroxide levels were measured by using the dye
2,7-dichlorofluorescein diacetate (DCF) as described previously
(Goodman and Mattson, 1994; Mattson et al., 1995). Ratiometric imaging
of the calcium indicator dye fura-2 was performed as described
previously (Mattson et al., 1992, 1993a). For measurements of DCF
fluorescence and [Ca2+]i after exposure to
A, cells were loaded with DCF or fura-2 and maintained in the
presence of A during imaging.
RESULTS
PC12 cells expressing mutant PS-1 exhibit increased vulnerability
to apoptosis induced by serum withdrawal and amyloid -peptide
Rat neural (PC12) cells were stably transfected with a DNA
construct encoding PS-1 containing the 286 leu-val mutation (L286V), and additional lines were transfected with wild-type PS-1 or vector alone. RT-PCR analysis showed that PS-1 and PS-1 L286V mRNAs were expressed at variable levels in each of the transfected cell lines isolated (data not shown) (cf. Guo et al., 1996). Levels of PS-1 expression were established in Western blot analyses (Fig.
1); clonal lines expressing moderately high levels of
wild-type PS-1 (n = 3) or PS1L286V (n = 3), one vector-transfected line, and the untransfected parent cell line
were chosen for use in the present study. Densitometric analyses of
Western blots showed that the levels of expression of wild-type and
mutant PS-1 in the clones chosen were very similar and were at least
five times greater than the background level of endogenous PS-1 (Fig.
1B). Under the normal culture maintenance conditions,
overexpression of wild-type or mutant PS-1 at the levels of the lines
used in the present study did not seem to affect cell survival or
growth.
Fig. 1.
Expression of wild-type and mutant PS-1 in
PC12 cells. A, Western blot showing PS-1 protein in
untransfected and vector-transfected PC12 cells and in three different
lines of cells expressing wild-type (PS-1) and mutant
(PS1L286V) PS-1. Equivalent amounts of protein (50 µg/lane) from cell homogenates of the indicated cell lines (untransfected cells, vector-transfected cells, PS-1, and PS1L286V) were separated by SDS-PAGE, transferred to nitrocellulose, and probed
with PS-1 antibody. Similar to results of other investigators (Thinakaran et al., 1996), in addition to recognizing full-length PS-1
(46 kDa band), our anti-loop PS-1 antibody also recognized presumptive
proteolytic products of PS-1 ~32 and 19 kDa. B,
Densitometric analysis of relative levels of PS-1 protein in PC12 cell
lines stably expressing wild-type (PS-1) and mutant
(PS1L286V) PS-1. Note that basal PS-1 levels are
relatively low in PC12 cells and that levels of expression of wild-type
and mutant PS-1 were similar among the lines shown.
[View Larger Version of this Image (26K GIF file)]
To determine whether the expression of PS-1 L286V affected cell death
after trophic factor withdrawal, we deprived various lines of
undifferentiated PC12 cells of serum, an insult previously shown to
induce apoptosis (Bastitatou and Greene, 1991; Rukenstein et al.,
1991). In untransfected and vector-transfected PC12 cells there was a
progressive appearance of cells with apoptotic nuclei that occurred
between 12 and 48 hr after serum withdrawal (Fig. 2A). In PC12 cells expressing PS-1
L286V there was a dramatic increase in the numbers of apoptotic cells
present at 12, 24, and 48 hr after serum withdrawal (Fig.
2A,C). In another set of experiments cells were
differentiated into a neuron-like phenotype by chronic exposure to NGF,
and then the NGF was withdrawn. Apoptosis induced by NGF withdrawal in
cell lines expressing mutant PS-1 was significantly greater than in
untransfected cells, vector-transfected cells, and cells overexpressing
wild-type PS-1 (Fig. 2B). NGF withdrawal induced
apoptosis in ~20% of cells in untransfected and vector-transfected
cell lines and in ~80% of the cells in lines expressing mutant PS-1.
The level of apoptosis after NGF withdrawal in cells overexpressing
wild-type PS-1 was somewhat higher than that in vector-transfected and
untransfected lines, although the difference did not reach statistical
significance (Fig. 2B).
Fig. 2.
PC12 cells expressing PS-1 L286V mutation exhibit
increased vulnerability to apoptosis induced by trophic factor
withdrawal. A, The percentages of cells exhibiting DNA
condensation and fragmentation (apoptotic cells) were determined in
Hoescht-stained cultures at the indicated times after serum withdrawal
from undifferentiated PC12 cell lines: untransf,
untransfected cells; Vector, cells transfected with
empty vector; PS-1, cells transfected with wild-type PS-1 (pooled data from three different lines; C1, C2, and C6); and
three different lines expressing mutant PS-1
(PS1L286V). Values represent the mean and SD of
determinations made in four separate cultures (200 cells
counted/culture). Values for each of the three cell lines expressing
L286V were greater than corresponding values for untransfected cultures
and cultures transfected with vector or wild-type PS-1 (12 hr time
point, p < 0.05; 24 and 48 hr time points,
p < 0.001); ANOVA with Scheffé's
post hoc tests. B, Control untransfected
PC12 cells (Control) and PC12 cells transfected with empty vector (Vector), wild-type PS-1
(PS-1), or mutant PS-1 (PS1L286V)
were differentiated in the presence of NGF (data pooled from analyses
on all three lines expressing wild-type PS-1 and all three lines
expressing PS-1 L286V; compare with Fig. 1). NGF was withdrawn, and 48 hr later the percentages of apoptotic cells in each culture were
determined. Values represent the mean and SD of determinations made in
four separate cultures. *p < 0.01 compared with
each of the other values; ANOVA with Scheffé's post
hoc tests. C, Confocal images of propidium
iodide fluorescence in vector-transfected and L286V-transfected PC12
cells 24 hr after serum withdrawal. Fluorescence intensity is depicted
in pseudocolor according to the color scale bar. Note
that DNA is distributed diffusely throughout the nuclei of most
vector-transfected cells, whereas DNA is fragmented to varying extents
in most cells expressing PS-1 L286V (arrowheads).
[View Larger Version of this Image (97K GIF file)]
Previous studies showed that A can induce apoptosis in cultured
primary neurons (Loo et al., 1993) and PC12 cells (Gschwind and Huber,
1995). Exposure of undifferentiated PC12 cells to A for 24 hr
induced apoptotic nuclear changes, and the percentage of cells
undergoing apoptosis was significantly greater in cells expressing PS-1
L286V than in untransfected cells, vector-transfected cells, and cells
overexpressing wild-type PS-1 (Fig. 3A).
Nuclear condensation and fragmentation induced by A (Fig.
3A) and serum withdrawal (data not shown) were prevented by
the protein synthesis inhibitor cycloheximide, consistent with an
apoptotic mechanism of cell death. It was reported previously that A
causes a relatively rapid (minutes to hours) impairment of
mitochondrial function as measured by MTT reduction that occurs very
early in the apoptotic process (Shearman et al., 1994; Kruman et al.,
1996). Exposure of PC12 cells to A resulted in a decrease in levels
of MTT reduction in all cell lines examined, and the magnitude of the
decrease in MTT reduction was significantly greater in each line
expressing PS-1 L286V than in untransfected cells, cells transfected
with empty vector, and lines overexpressing wild-type PS-1 (Fig.
3B).
Fig. 3.
Mutant PS-1 increases PC12 cell
vulnerability to apoptosis and mitochondrial dysfunction induced by
amyloid -peptide. A, The indicated cell lines were
exposed to vehicle (Vehicle), 50 µM A
(A), or 10 µM cycloheximide plus 50 µM A (CHX + A) for 24 hr. Then cells
were stained with Hoescht dye, and the percentages of cells exhibiting
DNA condensation and fragmentation were determined. Values represent
the mean and SD of determinations made in four separate cultures (data
pooled from analyses on all three lines expressing wild-type PS-1 and
all three lines expressing PS-1 L286V; compare with Fig. 1). For all
cell lines the values for cells exposed to A were significantly
greater than values for vehicle or CHX plus A-treated cell lines
(p < 0.01). *p < 0.01 compared with each of the other cell lines exposed to A; ANOVA with
Scheffé's post hoc tests. B,
Parallel cultures of untransfected control cells
(Untransf), vector-transfected cells
(Vector), three lines of cells transfected with
wild-type PS-1 (PS-1; pooled data), and three lines of
mutant PS-1 cells (PS1L286V; pooled data) were exposed
for 4 hr to 50 µM A, and relative levels of MTT
reduction (a measure of mitochondrial function) were quantified. Values are the mean and SD of determinations made in four separate cultures and are expressed as a percentage of vehicle-treated control
(vector-transfected) cells (data pooled from analyses on all three
lines expressing wild-type PS-1 and all three lines expressing PS-1
L286V; compare with Fig. 1). There were no differences in basal levels
of MTT reduction among the various control, wild-type PS-1-expressing, and mutant PS-1-expressing lines (data not shown).
*p < 0.01 compared with corresponding values for
untransfected, vector-transfected, and WT PS-1-transfected lines
exposed to A; ANOVA with Scheffé's post hoc
tests.
[View Larger Version of this Image (21K GIF file)]
Proapoptotic action of PS-1 mutation involves disruption of calcium
homeostasis and induction of oxidative stress
To test the hypothesis that PS-1 mutations promote cell death by
increasing oxidative stress, we determined whether antioxidants would
protect cells against death induced by A and measured levels of
peroxides in the different cell lines after exposure to A. When
cultures were pretreated with the antioxidants propyl gallate or
vitamin E before exposure to A, cell death 24 hr later was reduced
significantly, and the death-enhancing effect of L286V was prevented
(Fig. 4A). In light of previous data
implicating disruption of cellular calcium homeostasis in the mechanism
of A cytotoxicity (Mattson et al., 1992, 1993a; Mark et al., 1995) and apoptosis (Takei and Endo, 1994; Ciutat et al., 1995), we determined whether cells expressing PS-1 L286V exhibit increased sensitivity to A-induced elevation of
[Ca2+]i and whether agents that suppress
calcium influx would protect cells against the adverse effects of PS-1
L286V. A induced an increase in [Ca2+]i
during a 4 hr exposure period; the elevation of
[Ca2+]i was significantly greater in cells
expressing PS-1 L286V, as compared with control cell lines (Fig.
4B). A caused [Ca2+]i to
increase to 150-170 nM in control cell lines and to >300 nM in L286V-expressing cells. Cell death induced by A,
and the death-enhancing effect of PS-1 L286V was attenuated
significantly in cultures pretreated with nifedipine, a blocker of
L-type voltage-dependent calcium channels (Fig. 4A).
Dantrolene, an inhibitor of calcium release from ER stores, also in
large part prevented A-induced cell death in cells expressing L286V,
suggesting a role for altered calcium release in the proapoptotic
action of PS-1 L286V (Fig. 4A).
Nifedipine and dantrolene also afforded partial protection against the
adverse effects of mutant PS-1 on [Ca2+]i
(Fig. 4B) and cellular peroxide levels (Fig.
4C). As expected from previous studies (Behl et al., 1994;
Goodman and Mattson, 1994), exposure of PC12 cells to A for 4 hr
caused an increase in cellular peroxide levels; the increase was
significantly greater in cells expressing PS-1 L286V than in control
cell lines (Fig. 4C). Pretreatment with either propyl
gallate or vitamin E in large part abolished the A-induced increase
in peroxide levels (Fig. 4C). Collectively, these data
indicated that mutant PS-1 may sensitize neural cells to apoptosis by
perturbing calcium homeostasis and free radical metabolism.
Fig. 4.
PC12 cells expressing PS-1 L286V mutation exhibit
increased levels of oxidative stress and intracellular calcium after
exposure to A: attenuation by antioxidants and blockers of calcium
influx and release from intracellular stores. A,
Cultures were pretreated for 24 hr with 50 µM vitamin E
(VitE) or for 2 hr with 5 µM propyl gallate (PG), 1 µM nifedipine (Nifedipine), or 1 µM dantrolene (DTL). Then cultures were
exposed to 50 µM A for 24 hr, and the medium was
removed for LDH assay (cells exposed to A first undergo apoptosis,
followed by secondary necrosis, the latter being detected by LDH
release assay). Values are expressed as a percentage of the maximal LDH
release (mean and SD of 6-8 cultures); maximal LDH release was
determined in parallel cultures (of each cell line) subjected to
freeze-thaw. Values for each of the A-treated cell lines expressing
PS-1 L286V were significantly greater than each of the values for
control (untransfected) and vector-transfected cell lines
(p < 0.01) and for each of the values in
PS-1 L286V cultures pretreated with vitamin E, propyl gallate,
nifedipine, or dantrolene (p < 0.01 in each
case); ANOVA with Scheffé's post hoc tests.
B, C, Cultures were pretreated with
antioxidants or Ca2+ flux blockers as described for
A and then exposed to vehicle or A for 4 hr. Then the
relative [Ca2+]i (fura-2 imaging)
(B) and levels of peroxides (DCF
Fluorescence) (C) in individual cells were
quantified. Values are the mean and SD of determinations made in three
to four cultures (15-25 cells for [Ca2+]i
measurements and 40-60 cells/culture for DCF measurements). Values for
each of the A-treated cell lines expressing L286V were significantly
greater than each of the values in the vehicle-treated cultures
(p < 0.01), each of the values in the
cultures pretreated with vitamin E or propyl gallate
(p < 0.01), and each of the values for
L286V cells in the cultures pretreated with nifedipine or dantrolene
(p < 0.05); ANOVA with Scheffé's
post hoc tests.
[View Larger Version of this Image (28K GIF file)]
Apoptosis in differentiated PC12 cells induced by NGF withdrawal is
enhanced by mutant PS-1
The experiments described above used PC12 cell lines
stably expressing high levels of wild-type or mutant PS-1, a situation that could perturb metabolic pathways nonspecifically in the cells. Moreover, because wild-type and mutant PS-1 were
expressed throughout the period of NGF-induced cell differentiation, we
could not rule out the possibility that mutant PS-1 affects the
differentiation process. To examine further the proapoptotic actions of
PS-1 mutations, we therefore established PC12 lines expressing
wild-type and mutant PS-1 under the control of a
tetracycline-suppressible (Tet-off) promoter. For these experiments
cells were maintained in the presence of tetracycline during the period
of differentiation with NGF. After differentiation and 48 hr before NGF
withdrawal, tetracycline concentration was reduced to induce PS-1
wild-type or mutant expression. As expected, levels of PS-1 expression
increased with decreasing concentrations of tetracycline in the culture
medium (Fig. 5A); subsequent
experiments were performed in cultures induced to express wild-type or
mutant PS-1 at comparable levels (Fig. 5B). Withdrawal of
NGF from differentiated untransfected, vector-transfected, and
Tet-off-transfected PC12 cells resulted in apoptosis of ~40% of the
cells during a 48 hr period (Fig. 6A).
In contrast, apoptosis did not occur after NGF withdrawal in PC12 cells
expressing Bcl-2. NGF withdrawal-induced apoptosis was enhanced
significantly in PC12 cells expressing mutant PS-1 L286V (Fig.
6C), as compared with cells expressing wild-type PS-1 (Fig.
6B). Cells expressing PS-1 L286V also exhibited a
modest increase in basal levels of apoptosis in the presence of NGF.
PC12 cells expressing Bcl-2 in combination with wild-type or mutant
PS-1 were completely resistant to apoptosis induced by NGF withdrawal
(Fig. 6C). Collectively, these data indicate that mutant
PS-1 possesses an adverse property not present in wild-type PS-1 that
sensitizes neurons to apoptosis.
Fig. 5.
Controlled expression of PS-1 in PC12 cells with
the use of a tetracyline-responsive transactivator. A, A
PC12 cell line expressing the "Tet-off" construct was stably
transfected with a pTRE-derived plasmid expressing PS-1 L286V gene.
Cells were incubated for 48 hr in the presence of 2.0, 0.004, 0.002, and 0 µg/ml tetracycline (lanes 1-4,
respectively). Cell proteins were separated by SDS-PAGE (100 µg/lane), transferred to a nitrocellulose sheet, and
immunoreacted with PS-1 antibody. Note that, as the concentration of
tetracycline was decreased, the levels of mutant PS-1 expression
increased. B, Western blot showing that in the absence
of tetracycline a double-stable PS-1 L286V cell line (C1) and a
double-stable PS-1 cell line (C3) show significantly higher levels of
PS-1 expression than do vector-transfected or untransfected cell
lines.
[View Larger Version of this Image (43K GIF file)]
Fig. 6.
Mutant PS-1 increases the vulnerability of
differentiated PC12 cells to NGF withdrawal-induced apoptosis. Cultures
of differentiated PC12 cells were incubated for 48 hr in serum-free
medium containing or lacking NGF, and the percentage of cells
exhibiting nuclear condensation and fragmentation was quantified.
Values are the mean and SEM of determinations made in at least four
separate cultures. A, Analyses in various control PC12
cell lines: WT, untransfected wild-type cells;
puro, vector-transfected cells (pBabe-puro vector used
for Bcl-2 expression); Tet-off, cells transfected with
the Tet-off plasmid; puro Tet-off, cells doubly transfected with pBabe-puro vector and Tet-off plasmid; bcl-2 puro, cells expressing Bcl-2. Note that NGF withdrawal induced a similar level of apoptosis in all control lines, whereas cells expressing Bcl-2 were resistant to NGF withdrawal-induced apoptosis. *p < 0.01 compared with corresponding values for
NGF+ cultures and the value for Bcl-2 NGF
cells. B, Bcl-2 protects PC12 cells overexpressing
wild-type PS-1 against NGF withdrawal-induced apoptosis. Two lines of
control cells expressing wild-type PS-1 (C3 and
C7) and two different lines of cells expressing
both Bcl-2 and PS-1 (C11 and C13) were analyzed. *p < 0.01 compared with corresponding
values for NGF+ cells and compared with the value for the
NGF line expressing Bcl-2. C, Mutant PS-1
enhances vulnerability of PC12 cells to apoptosis induced by NGF
withdrawal: protection by Bcl-2. Two lines of control cells expressing
PS-1 L286V and two lines of cells expressing both Bcl-2 and PS-1 L286V
were analyzed. *p < 0.05 compared with
corresponding values for each line expressing wild-type PS-1
(B), p < 0.01 compared with
corresponding values for cells maintained in the presence of NGF, and
p < 0.01 compared with NGF
lines coexpressing Bcl-2; ANOVA with Scheffé's post
hoc tests for pair-wise comparisons.
[View Larger Version of this Image (18K GIF file)]
Enhanced oxidative stress after NGF withdrawal and perturbed
calcium homeostasis in differentiated PC12 cells expressing mutant
PS-1
Withdrawal of NGF from differentiated untransfected and
vector-transfected PC12 cells resulted in an increase in levels of cellular peroxides that occurred within 3 hr (Fig.
7A). An increase in cellular peroxides also
occurred after NGF withdrawal from PC12 cells expressing Bcl-2,
suggesting that the antiapoptotic action of Bcl-2 occurred downstream
of the oxidative stress (cf. Greenlund et al., 1995). NGF
withdrawal-induced peroxide accumulation was enhanced greatly in PC12
cells expressing the mutant PS-1 L286V, as compared with cells
expressing wild-type PS-1 (Fig. 7B). Basal levels of
peroxides in cells expressing PS-1 L286V were somewhat higher than in
control cells, although the difference did not reach statistical
significance. PC12 cells expressing Bcl-2 in combination with mutant
PS-1 exhibited peroxide levels after NGF withdrawal that were lower
than in cells lacking Bcl-2, but the difference did not reach
statistical significance (Fig. 7C).
Fig. 7.
Oxidative stress induced by NGF withdrawal is
enhanced in PC12 cells expressing mutant PS-1. Cultures of
differentiated PC12 cells were incubated for 3 hr in serum-free medium
containing or lacking NGF, and levels of cellular peroxides were
quantified by confocal laser scanning microscope image analysis of DCF
fluorescence. Values are the mean and SEM of determinations made in at
least four separate cultures. A, Analyses in various
control PC12 cell lines: WT, untransfected wild-type
cells; puro, vector-transfected cells;
Tet-off, cells transfected with the Tet-off plasmid;
puro Tet-off, cells doubly transfected with empty vector
and Tet-off plasmid; bcl-2 puro, cells expressing Bcl-2.
Note that NGF withdrawal induced a similar level of peroxide
accumulation in all control lines and in cells overexpressing Bcl-2.
Each value for NGF cultures was significantly greater
than the corresponding value for NGF+ cells
(p < 0.01). B, Bcl-2 does
not prevent NGF withdrawal-induced accumulation of peroxides in PC12
cells overexpressing wild-type PS-1. Two lines of control cells
expressing wild-type PS-1 (C3 and
C7) and two different lines of cells expressing
both Bcl-2 and PS-1 (C11 and C13) were
analyzed. Each value for NGF
cultures was significantly greater than
the corresponding value for NGF+ cells
(p < 0.01). C, Mutant PS-1
enhances accumulation of peroxides in PC12 cells deprived of NGF. Two
lines of control cells expressing PS-1 L286V and two lines of cells
expressing both Bcl-2 and PS-1 L286V were analyzed.
*p < 0.05 compared with corresponding values for
each line expressing wild-type PS-1 (B) and
p < 0.01 compared with corresponding values for
cells maintained in the presence of NGF; ANOVA with Scheffé's
post hoc tests for pair-wise comparisons).
[View Larger Version of this Image (20K GIF file)]
PS-1 is localized to the ER, and a recent study showed that PC12
cells expressing mutant PS-1 exhibit altered ER calcium regulation (Guo
et al., 1996). We therefore examined calcium responses to thapsigargin,
an inhibitor of the ER Ca2+-ATPase in differentiated PC12
cells expressing wild-type or mutant PS-1, with or without Bcl-2. Rest
[Ca2+]i was ~60 nM in control
cells and was elevated to 80-110 nM in cells expressing
PS-1 L286V (Fig. 8). In PC12 cells not expressing PS-1,
thapsigargin caused an increase in [Ca2+]i to
160-180 nM, and the response was attenuated by ~50% in
cells expressing Bcl-2 (Fig. 8A). PC12 cells
expressing PS-1 L286V exhibited a markedly en- hanced peak
[Ca2+]i response to thapsigargin, with levels
rising to ~400 nM (Fig. 8C). The enhanced
[Ca2+]i response to thapsigargin was
abolished completely in PC12 cells expressing Bcl-2 (Fig.
8C). Taken together with the data above showing that
dantrolene protects PC12 cells against the proapoptotic actions of PS-1
L286V, these findings suggest that altered ER calcium regulation is
involved mechanistically in the apoptotic action of mutant PS-1.
Fig. 8.
Elevations of
[Ca2+]i induced by thapsigargin are enhanced
significantly in PC12 cells expressing mutant PS-1: attenuation by Bcl-2. PC12 cells were incubated in serum-free medium and basal [Ca2+]i (Tpg),
and the peak [Ca2+]i after exposure to 1 µM thapsigargin (Tpg+) was
quantified (cf. Guo et al., 1996). Values are the mean and SEM of
determinations made in at least four separate cultures (15-20
cells/culture). A, Analyses in various control PC12 cell lines: WT, untransfected wild-type cells;
puro, vector-transfected cells (vector for
Bcl-2-expressing line); Tet-off, cells transfected with
the Tet-off plasmid; puro Tet-off, cells doubly
transfected with pBabe-puro and Tet-off plasmid; bcl-2
puro, cells expressing Bcl-2. *p < 0.01 compared with corresponding values for Tpg cultures, and
p < 0.05 compared with the Tpg+ value
in cells expressing Bcl-2. B, The
[Ca2+]i response to thapsigargin in cells
expressing wild-type PS-1 is attenuated in cells coexpressing Bcl-2.
*p < 0.01 compared with corresponding values for
Tpg, and p < 0.05 compared with
Tpg+ values in cells expressing Bcl-2. C,
Mutant PS-1 enhances [Ca2+]i responses to
thapsigargin: attenuation by Bcl-2. *p < 0.001 compared with corresponding values for Tpg,
p < 0.01 compared with Tpg+ values in
cells expressing Bcl-2, and p < 0.01 compared with Tpg+- PS1 Tet-off values (B); ANOVA with
Scheffé's post hoc tests for pair-wise
comparisons.
[View Larger Version of this Image (19K GIF file)]
DISCUSSION
Previous studies showed that both A (Rabizadeh et al., 1994;
Gschwind and Huber, 1995) and trophic factor withdrawal (Bastitatou and
Greene, 1991; Rukenstein et al., 1991) induce apoptosis in PC12 cells.
Similarly, A (Forloni et al., 1993; Loo et al., 1993) and trophic
factor deprivation (Prehn et al., 1994) induce apoptosis in neurons in
primary cultures established from brain regions (e.g., hippocampus,
neocortex, and basal forebrain) that are affected in AD. We found that
both undifferentiated and differentiated PC12 cells expressing PS-1
L286V were extremely sensitive to apoptotic cell death when compared
with various control cell lines. These findings suggest that the
mutated PS-1 protein possesses an adverse proapoptotic property.
Overexpression of wild-type PS-1, at levels similar to or greater than
mutant PS-1 levels, did not result in increased vulnerability of PC12
cells to apoptosis, indicating that the proapoptotic action of PS-1
L286V was not simply the consequence of increased levels of PS-1
protein. Although the specific nature of that adverse property of the
PS-1 mutation was not established in the present study, the data
suggest an action on systems that regulate free radical metabolism
and/or calcium homeostasis. Thus, levels of cellular peroxides induced by A were increased greatly in cells expressing the PS-1 mutation, as compared with control lines, and two different antioxidants (vitamin
E and propyl gallate) protected PC12 cells against cell death induced
by A. The antioxidants also suppressed A-induced increases in
intracellular peroxide and calcium levels, consistent with previous
data suggesting that the mechanism of A neurotoxicity involves
membrane lipid peroxidation and impairment of membrane ion transport
systems and calcium influx (Mattson et al., 1992; Behl et al., 1994;
Goodman and Mattson, 1994; Mark et al., 1995, 1997a,b).
Previous studies of mechanisms of neuron death induced by trophic
factor withdrawal and A have implicated reactive oxygen species
(ROS) (Hockenbery et al., 1993; Kane et al., 1993; Greenlund et al.,
1995). We found that levels of oxidative stress and apoptosis after NGF
withdrawal from differentiated PC12 cells were enhanced in cells
expressing mutant PS-1, but not in cells overexpressing wild-type PS-1.
These effects of mutant PS-1 were not attributable to changes that
occurred during the process of cellular differentiation, because we
allowed the cells to differentiate before induction of PS-1 expression
by using the Tet-off system. Considerable data indicate that levels of
oxidative stress are increased in AD brain, particularly in the
environment of neuritic plaques and in neurofibrillary tangles (for
review, see Benzi and Moretti, 1995; Smith et al., 1995);
levels of oxidative stress also are increased in the brain during
normal aging (Stadtman, 1992). The present findings suggest the
possibility that PS-1 mutations may promote oxidative stress and
thereby sensitize neurons to decrements in trophic factor support and
increased accumulations of A that occur in the aging brain. The data
also may provide at least a partial explanation for the increased
production of A documented in blood and other tissues from human
carriers of PS-1 mutations (Scheuner et al., 1996), because studies
have shown that manipulations that promote metabolic stress and
increase [Ca2+]i in cultured neurons can
alter proteolytic processing of -amyloid precursor protein (APP)
in cultured cells in favor of increased A production (Gabuzda et
al., 1994; Querfurth and Selkoe, 1994).
PS-1 seems to be localized to the ER in several cell types, including
neurons (Guo et al., 1996; Kovacs et al., 1996; Walter et al., 1996).
Calcium imaging studies of cultured PC12 expressing PS-1 L286V have
shown that this mutation alters calcium release from ER stores such
that calcium responses to agonists that activate the IP3
pathway (e.g., muscarinic cholinergic agonists and bradykinin) are
enhanced greatly (Guo et al., 1996). The perturbed calcium homeostasis
observed in PC12 cells expressing mutant PS-1 is consistent with
reports that calcium signaling is altered in cultured fibroblasts taken
from carriers of PS-1 mutations (McCoy et al., 1993; Ito et al., 1994).
Our data suggest that disruption of calcium homeostasis by mutant PS-1
could be linked mechanistically to its proapoptotic action because
dantrolene, an agent that blocks calcium release from ER, protected
cells against the death-promoting effect of the PS-1 mutation. Recent
findings in studies of non-neuronal cells have linked ER calcium
regulation to apoptosis. For example, Lam et al. (1993) showed that
glucocorticoids induce release of calcium from ER, which is correlated
with subsequent DNA fragmentation and apoptosis, and Khan et al. (1996)
provided evidence that lymphocyte apoptosis is mediated by increased
expression of IP3 receptors. We found that
thapsigargin-induced increases of [Ca2+]i
were enhanced in PC12 cells expressing mutant PS-1 and that Bcl-2
prevented the enhanced response in cells expressing mutant PS-1. Lam et
al. (1994) showed that Bcl-2 protects lymphoma cells against apoptosis
induced by thapsigargin and suppresses calcium release from ER. The
localization of mutant PS-1 to ER (Guo et al., 1996) suggests that its
effects on ER calcium homeostasis and apoptosis may be linked
mechanistically, a possibility supported by our data. The ability of
dantrolene to suppress the increased peroxide accumulation induced by
A in PC12 cells expressing PS-1 L286V suggests that calcium release
from ER contributes to the enhanced oxidative stress associated with
mutant PS-1 expression. However, in addition to calcium release from
ER, calcium influx through voltage-dependent plasma membrane channels
also may be involved, because nifedipine protected cells against A
toxicity and suppressed elevation of [Ca2+]i
and peroxide accumulation induced by A. These data are consistent with previous studies of primary neurons and PC12 cells showing that
removal of extracellular calcium or treatment with nifedipine attenuates A toxicity (Mattson et al., 1993a; Weiss et al., 1994).
The observation that both antioxidants and calcium channel blockers
protected neurons against the adverse effects of the PS-1 mutation are
consistent with a scenario in which A induces a vicious cycle in
which oxidative stress disrupts ion homeostasis, which, in turn,
promotes further oxidative stress. There is now abundant evidence to
support the involvement of such reciprocating cytotoxic cascades in
many different neurodegenerative conditions (Mattson et al., 1992;
Zhang et al., 1993; Goodman et al., 1996). Thus, although both the
normal function of PS-1 and the exact alteration that results from PS-1
mutations are unknown, our data indicate that PS-1 mutations promote
neurodegenerative apoptotic cascades involving perturbed ion
homeostasis and oxidative stress.
Recent data from studies of brain tissue, blood, and cultured
fibroblasts from carriers of PS mutations and studies of transfected cell lines indicate that cells expressing PS mutations produce greater
than normal levels of A1-42 (Borchelt et al., 1996; Lemere et al.,
1996; Scheuner et al., 1996). In addition, studies of transgenic mice
that express mutant PS-1 suggest that levels of A1-42 are increased
in brain tissue (Duff et al., 1996). The latter data suggest that PS-1
mutations may cause early onset AD by altering APP processing in
ways that lead to increased A production. The increased
vulnerability of neuronal cells expressing PS-1 L286V to A toxicity
and trophic factor withdrawal documented in the present study is
unlikely to result from increased A production because PC12 cells
are a rat cell line and, in contrast to human A, rat A is neither
amyloidogenic nor neurotoxic (Otvos et al., 1993). However, it is
conceivable that altered processing of APP could reduce levels of
neuroprotective secreted forms of APP, which have been shown to protect
neurons against oxidative apoptotic insults, including A toxicity
and glucose withdrawal (Mattson et al., 1993b; Goodman and Mattson,
1994; Furukawa et al., 1996).
Collectively, our data suggest that mutant PS-1 protein possesses an
adverse proapoptotic property. Wolozin et al. (1996) recently reported
that PC12 cells overexpressing wild-type PS-2 exhibit increased
apoptosis after trophic factor withdrawal and that a PS-2 mutant
enhanced basal levels of apoptosis. Deng et al. (1996) reported that
overexpression of wild-type PS-2 increased vulnerability of PC12 cells
to apoptosis induced by staurosporine or hydrogen peroxide. ALG-3, the
mouse homolog of PS-2, was shown to modulate apoptosis in T
lymphocytes (Vito et al., 1996), although in the latter case ALG-3
prevented apoptosis. We observed neither increased nor reduced
apoptosis in PC12 cells overexpressing wild-type PS-1, suggesting that
PS-1 does not function directly in an apoptotic pathway and that mutant
PS-1 acquires a novel adverse property. However, we cannot rule
out the possibility that mutant PS-1 interferes with a normal trophic
property of endogenous PS-1 in a loss-of-function scenario.
FOOTNOTES
Received Feb. 12, 1997; revised March 20, 1997; accepted March 25, 1997.
This work was supported by Grants to M.P.M. from National Institutes of
Health (NS30583 and AG10836) and the Alzheimer's Association, to
G.M.M. from National Institutes of Health (AG10917), and to B.L.S. from
the University of Washington Nathan Shock Center for Excellence in the
Basic Biology of Aging. We thank J. Begley, S. Bose, R. Pelfrey, and J. Xie for technical assistance.
Correspondence should be addressed to Dr. Mark P. Mattson, 211 Sanders-Brown Building, University of Kentucky, Lexington, KY
40536-0230.
REFERENCES
-
Bastitatou A,
Greene LA
(1991)
Aurintricarboxylic acid rescues PC12 cells and sympathetic neurons from cell death caused by nerve growth factor deprivation: correlation with suppression of endonuclease activity.
J Cell Biol
115:461-471[Abstract/Free Full Text].
-
Behl C,
Davis JB,
Lesley R,
Schubert D
(1994)
Hydrogen peroxide mediates amyloid -protein toxicity.
Cell
77:817-827[Web of Science][Medline].
-
Benzi G,
Moretti A
(1995)
Are reactive oxygen species involved in Alzheimer's disease?
Neurobiol Aging
16:661-674[Web of Science][Medline].
-
Black MM,
Greene LA
(1982)
Changes in the colchicine susceptibility of microtubules associated with neurite outgrowth: studies with nerve growth factor-responsive PC12 pheochromocytoma cells.
J Cell Biol
95:379-386[Abstract/Free Full Text].
-
Borchelt DR,
Thinakaran G,
Eckman CB,
Lee MK,
Davenport F,
Ratovitsky T,
Prada C-M,
Kim G,
Seekins S,
Yager D,
Slunt HH,
Wang R,
Seeger M,
Levey AI,
Gandy SE,
Copeland NG,
Jenkins NA,
Price DL,
Younkin SG,
Sisodia SS
(1996)
Familial Alzheimer's disease-linked presenilin-1 variants elevate A1-42/1-40 ratio in vitro and in vivo.
Neuron
17:1005-1013[Web of Science][Medline].
-
Bredesen DE
(1995)
Neural apoptosis.
Ann Neurol
38:839-851[Web of Science][Medline].
-
Bruce AJ,
Malfroy B,
Baudry M
(1996)
-amyloid toxicity in organotypic cultures: protection by EUK-8, a synthetic catalytic free radical scavenger.
Proc Natl Acad Sci USA
88:3633-3636[Abstract/Free Full Text].
-
Ciutat D,
Esquerda JE,
Caldero J
(1995)
Evidence for calcium regulation of spinal cord motoneuron death in the chick embryo in vivo.
Dev Brain Res
86:167-179[Medline].
-
Cook DB,
Sung JC,
Golde TE,
Felsenstein KM,
Wojczyk BS,
Tanzi RE,
Trojanowski JQ,
Lee VMY,
Doms RW
(1996)
Expression and analysis of presenilin-1 in a human neuronal system: localization in cell bodies and dendrites.
Proc Natl Acad Sci USA
93:9223-9228[Abstract/Free Full Text].
-
Cribbs DH,
Chen L,
Bendle SM,
La Ferla FM
(1996)
Widespread neuronal expression of the presenilin-1 early-onset Alzheimer's disease in the murine brain.
Am J Pathol
148:1797-1806[Abstract].
-
Deng G,
Pike CJ,
Cotman CW
(1996)
Alzheimer-associated presenilin-2 confers increased sensitivity to apoptosis in PC12 cells.
FEBS Lett
397:50-54[Web of Science][Medline].
-
Doan A,
Thinakaran G,
Borchelt DR,
Slunt HH,
Ratovitsky T,
Podlisny M,
Selkoe DJ,
Seeger M,
Gandy SE,
Price DL,
Sisodia SS
(1996)
Protein topology of presenilin-1.
Neuron
17:1023-1030[Web of Science][Medline].
-
Duff K,
Eckman C,
Zehr C,
Yu X,
Prada C-M,
Perez-Tur J,
Hutton M,
Buee L,
Harigaya Y,
Yager D,
Morgan D,
Gordon MN,
Holcomb L,
Refolo L,
Zenk B,
Hardy J,
Younkin S
(1996)
Increased amyloid-42(43) in brains of mice expressing mutant presenilin-1.
Nature
383:710-713[Medline].
-
Elder GA,
Tezapsidis N,
Carter J,
Shioi J,
Bouras C,
Li D,
Johnston JM,
Efthimiopoulos S,
Friedrich Jr VL,
Robakis NK
(1996)
Identification and neuron-specific expression of the S182/presenilin-1 protein in human and rodent brains.
J Neurosci Res
45:308-320[Web of Science][Medline].
-
Forloni G,
Chiesa R,
Smiroldo S,
Verga L
(1993)
Apoptosis-mediated neurotoxicity induced by chronic application of -amyloid fragment 25-35.
NeuroReport
4:523-526[Web of Science][Medline].
-
Fukuyama R,
Wadhwani KC,
Galdzicki Z,
Rapoport SI,
Ehrenstein G
(1994)
-Amyloid polypeptide increases calcium uptake in PC12 cells: a possible mechanism for its cellular toxicity in Alzheimer's disease.
Brain Res
667:269-272[Web of Science][Medline].
-
Furukawa K,
Sopher B,
Rydel RE,
Begley JG,
Martin GM,
Mattson MP
(1996)
Increased activity-regulating and neuroprotective efficacy of
-secretase-derived secreted APP is conferred by a C-terminal heparin-binding domain.
J Neurochem
67:1882-1896[Web of Science][Medline]. -
Gabuzda D,
Busciglio J,
Chen L,
Matsudaira P,
Yankner BA
(1994)
Inhibition of energy metabolism alters the processing of amyloid precursor protein and induces a potentially amyloidogenic derivative.
J Biol Chem
269:13623-13628[Abstract/Free Full Text].
-
Games D,
Adams D,
Alessandrinl R,
Barbour R,
Berthelette P,
Blackwell C,
Carr T,
Clemens J,
Donaldson T,
Gillespie F,
Guido T,
Hagoplan S,
Johnson-Wood K,
Khan K,
Lee M,
Lelbowitz E,
McConlogue S,
Montoya-Zavala M,
Mucke L,
Paganini L,
Penniman E,
Power M,
Schenk D,
Seubert P,
Snyder B,
Soriano F,
Tan H,
Vitale J,
Wadsworth S,
Wolozin B,
Zhao J
(1995)
Alzheimer-type neuropathology in transgenic mice overexpressing V717F -amyloid precursor protein.
Nature
373:523-527[Medline].
-
Giannakopoulos P,
Bouras C,
Kovari E,
Shioi J,
Tezapsidis N,
Hof PR,
Robakis NK
(1997)
Presenilin-1 immunoreactive neurons are preserved in late-onset Alzheimer's disease.
Am J Pathol
150:429-436[Abstract].
-
Goodman Y,
Mattson MP
(1994)
Secreted forms of -amyloid precursor protein protect hippocampal neurons against amyloid -peptide-induce oxidative injury.
Exp Neurol
128:1-12[Web of Science][Medline].
-
Goodman Y,
Bruce AJ,
Cheng B,
Mattson MP
(1996)
Estrogens attenuate and corticosterone exacerbates excitotoxicity, oxidative injury, and amyloid -peptide toxicity in hippocampal neurons.
J Neurochem
66:1836-1844[Web of Science][Medline].
-
Greenlund LJ,
Deckwerth TL,
Johnson EM
(1995)
Superoxide dismutase delays neuronal apoptosis: a role for reactive oxygen species in programmed neuronal death.
Neuron
14:303-315[Web of Science][Medline].
-
Gschwind M,
Huber G
(1995)
Apoptotic cell death induced by -amyloid 1-42 peptide is cell type-dependent.
J Neurochem
65:292-300[Web of Science][Medline].
-
Guo Q,
Furukawa K,
Sopher BL,
Pham DG,
Robinson N,
Martin GM,
Mattson MP
(1996)
Alzheimer's PS-1 mutation perturbs calcium homeostasis and sensitizes PC12 cells to death induced by amyloid -peptide.
NeuroReport
8:379-383[Web of Science][Medline].
-
Hockenbery D,
Oltvai ZN,
Yin XM,
Milliman CL,
Korsmeyer SH
(1993)
Bcl-2 functions in an antioxidant pathway to prevent apoptosis.
Cell
75:241-251[Web of Science][Medline].
-
Hsaio K,
Chapman P,
Nilsen S,
Eckman C,
Harigaya Y,
Younkin S,
Yang F,
Cole G
(1996)
Correlative memory deficits, A elevation, and amyloid plaques in transgenic mice.
Science
274:99-103[Abstract/Free Full Text].
-
Ito E,
Oka K,
Etcheberrigaray R,
Nelson TJ,
McPhie DL,
Tofel-Grehl B,
Gibson GE,
Alkon DL
(1994)
Internal Ca2+ mobilization is altered in fibroblasts from patients with Alzheimer disease.
Proc Natl Acad Sci USA
91:534-538[Abstract/Free Full Text].
-
Kane DJ,
Sarafian TA,
Anton R,
Hahn H,
Gralla EB,
Valentine JS,
Ord T,
Bredesen DE
(1993)
Bcl-2 inhibition of neural death: decreased generation of reactive oxygen species.
Science
262:1274-1277[Abstract/Free Full Text].
-
Kovacs DM,
Fausett HJ,
Page KJ,
Kim T-W,
Moir RD,
Merriam DE,
Hollister RD,
Hallmark OG,
Mancini R,
Felsenstein KM,
Hyman BT,
Tanzi RE,
Wasco W
(1996)
Alzheimer-associated presenilins 1 and 2: neuronal expression in brain and localization to intracellular membranes in mammalian cells.
Nat Med
2:224-229[Web of Science][Medline].
-
Kruman I,
Guo Q,
Bruce AJ,
Bredesen DE,
Mattson MP
(1996)
Hydroxynonenal may mediate apoptotic neuronal death induced by trophic factor withdrawal and oxidative insults.
Soc Neurosci Abstr
22:1481.
-
Lam M,
Dubyak G,
Distelhorst CW
(1993)
Effect of glucocorticosteroid treatment on intracellular calcium homeostasis in mouse lymphoma cells.
Mol Endocrinol
7:686-693[Abstract/Free Full Text].
-
Lemere CA,
Lopera F,
Kosik KS,
Lendon CL,
Ossa J,
Saido TC,
Yamaguchi H,
Ruiz A,
Martinez A,
Madrigal L,
Hincapie L,
Arango JC,
Anthony DC,
Koo EH,
Goate AM,
Selkoe DJ,
Arango VJC
(1996)
The E280A presenilin-1 Alzheimer mutation produces increased A42 deposition and severe cerebellar pathology.
Nat Med
2:1146-1150[Web of Science][Medline].
-
Levy-Lahad E,
Wasco W,
Poorkaj P,
Romano DM,
Oshima J,
Pettingell WH,
Yu C-E,
Jondro PD,
Schmidt SD,
Wang K,
Crowley AC,
Fu Y-H,
Guenette SY,
Galas D,
Nemens E,
Wijsman EM,
Bird TD,
Schellenberg GD,
Tanzi RE
(1995)
Candidate gene for the chromosome 1 familial Alzheimer's disease locus.
Science
269:973-977[Abstract/Free Full Text].
-
Li X,
Greenwald I
(1996)
Membrane topology of the C. elegans SEL-12 presenilin.
Neuron
17:1015-1021[Web of Science][Medline].
-
Loo DT,
Copani A,
Pike CJ,
Whittemore ER,
Walencewicz AJ,
Cotman CW
(1993)
Apoptosis is induced by -amyloid in cultured central nervous system neurons.
Proc Natl Acad Sci USA
90:7951-7955[Abstract/Free Full Text].
-
Mark RJ,
Hensley K,
Butterfield DA,
Mattson MP
(1995)
Amyloid -peptide impairs ion-motive ATPase activities: evidence for a role in loss of neuronal Ca2+ homeostasis and cell death.
J Neurosci
15:6239-6249[Abstract].
-
Mark RJ,
Lovell MA,
Markesbery WR,
Uchida K,
Mattson MP
(1997a)
A role for 4-hydroxynonenal in disruption of ion homeostasis and neuronal death induced by amyloid -peptide.
J Neurochem
68:255-264[Web of Science][Medline].
-
Mark RJ,
Pang Z,
Geddes JW,
Mattson MP
(1997b)
Amyloid -peptide impairs glucose uptake in hippocampal and cortical neurons: involvement of membrane lipid peroxidation.
J Neurosci
17:1046-1054[Abstract/Free Full Text].
-
Mattson MP,
Cheng B,
Davis D,
Bryant K,
Lieberburg I,
Rydel RE
(1992)
-Amyloid peptides destabilize calcium homeostasis and render human cortical neurons vulnerable to excitotoxicity.
J Neurosci
12:376-389[Abstract].
-
Mattson MP,
Tomaselli K,
Rydel RE
(1993a)
Calcium-destabilizing and neurodegenerative effects of aggregated -amyloid peptide are attenuated by basic FGF.
Brain Res
621:35-49[Web of Science][Medline].
-
Mattson MP,
Cheng B,
Culwell A,
Esch F,
Lieberburg I,
Rydel RE
(1993b)
Evidence for excitoprotective and intraneuronal calcium-regulating roles for secreted forms of -amyloid precursor protein.
Neuron
10:243-254[Web of Science][Medline].
-
Mattson MP,
Barger SW,
Begley JG,
Mark RJ
(1995)
Calcium, free radicals, and excitotoxic neuronal death in primary cell culture.
Methods Cell Biol
46:187-216[Web of Science][Medline].
-
McCoy KR,
Mullins RD,
Newcomb TG,
Ng GM,
Pavlinkova G,
Polinsky RJ,
Nee LE,
Sisken JE
(1993)
Serum- and bradykinin-induced calcium transients in familial Alzheimer's fibroblasts.
Neurobiol Aging
14:447-455[Web of Science][Medline].
-
Mullan M,
Crawford F
(1993)
Genetic and molecular advances in Alzheimer's disease.
Trends Neurosci
16:398-403[Web of Science][Medline].
-
Murphy GM,
Forno LS,
Ellis WG,
Nochlin D,
Levy-Lahad E,
Poorkaj P,
Bird TD,
Jiang Z,
Cordell B
(1996)
Antibodies to presenilin proteins detect neurofibrillary tangles in Alzheimer's disease.
Am J Pathol
149:1839-1846[Abstract].
-
Otvos L,
Szendrei GI,
Lee VM,
Mantsch HH
(1993)
Human and rodent Alzheimer -amyloid peptides acquire distinct conformations in membrane-mimicking solvents.
Eur J Biochem
211:249-257[Web of Science][Medline].
-
Prehn JH,
Bindokas VP,
Marcuccilli CJ,
Krajewski S,
Reed JC,
Miller RJ
(1994)
Regulation of neuronal Bcl2 protein expression and calcium homeostasis by transforming growth factor type beta confers wide-ranging protection on rat hippocampal neurons.
Proc Natl Acad Sci USA
91:12599-12603[Abstract/Free Full Text].
-
Querfurth HW,
Selkoe DJ
(1994)
Calcium ionophore increases amyloid -peptide production by cultured cells.
Biochemistry
33:4550-4561[Medline].
-
Rabizadeh S,
Bitler CM,
Butcher LL,
Bredesen DE
(1994)
Expression of the low-affinity nerve growth factor receptor enhances -amyloid peptide toxicity.
Proc Natl Acad Sci USA
91:10703-10706[Abstract/Free Full Text].
-
Rogaev EI,
Sherrington R,
Rogaeva EA,
Levesque G,
Ikeda M,
Liang Y,
Chi H,
Lin C,
Holman K,
Tsuda T,
Mar L,
Sorbi S,
Nacmias B,
Piacentini S,
Amaducci L,
Chumakov I,
Cohen D,
Lannfelt L,
Fraser PE,
Rommens JM,
St. George-Hyslop PH
(1995)
Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene.
Nature
376:775-778[Medline].
-
Rukenstein A,
Rydel RE,
Greene LA
(1991)
Multiple agents rescue PC12 cells from serum-free cell death by translation- and transcription-independent mechanisms.
J Neurosci
11:2552-2563[Abstract].
-
Scheuner D,
Eckman C,
Jensen M,
Song X,
Citron M,
Suzuki N,
Bird TD,
Hardy J,
Hutton M,
Kukull W,
Larson E,
Levy-Lahad E,
Viitanen M,
Peskind E,
Poorkaj P,
Schellenberg G,
Tanzi R,
Wasco W,
Lannfelt L,
Selkoe D,
Younkin S
(1996)
The amyloid -protein deposited in the senile plaques of Alzheimer's disease is increased in vivo by the presenilin 1 and 2 and APP mutations linked to familial Alzheimer's disease.
Nat Med
2:864-870[Web of Science][Medline].
-
Selkoe DJ
(1989)
Biochemistry of altered brain proteins in Alzheimer's disease.
Annu Rev Neurosci
12:463-490[Web of Science][Medline].
-
Shearman MS,
Ragan CI,
Iversen LL
(1994)
Inhibition of PC12 cell redox activity is a specific, early indicator of the mechanism of -amyloid-mediated cell death.
Proc Natl Acad Sci USA
91:1470-1474[Abstract/Free Full Text].
-
Sherrington R,
Rogaev EI,
Liang Y,
Rogaeva EA,
Levesque G,
Ikeda M,
Chi H,
Lin C,
Li G,
Holman K,
Tsuda T,
Mar L,
Foncin J-F,
Bruni AC,
Montesi MP,
Sorbi S,
Rainero I,
Pinessi L,
Nee L,
Chumakov I,
Pollen D,
Brookes A,
Sansequ P,
Polinsky RJ,
Wasco W,
Da Silva HAR,
Haines JL,
Pericak-Vance MA,
Tanzi RE,
Roses AD,
Fraser PE,
Rommens JM,
St. George-Hyslop PH
(1995)
Cloning of a gene bearing missense mutations in early-onset familial Alzheimer's disease.
Nature
375:754-760[Medline].
-
Smale G,
Nichols NR,
Brady DR
(1995)
Evidence for apoptotic cell death in Alzheimer's disease.
Exp Neurol
133:225-230[Web of Science][Medline].
-
Smith MA,
Sayre LM,
Monnier VM,
Perry G
(1995)
Radical aging in Alzheimer's disease.
Trends Neurosci
18:172-176[Web of Science][Medline].
-
Stadtman E
(1992)
Protein oxidation and aging.
Science
257:1220-1224[Abstract/Free Full Text].
-
Su JH,
Anderson AJ,
Cummings B,
Cotman CW
(1994)
Immunocytochemical evidence for apoptosis in Alzheimer's disease.
NeuroReport
5:2529-2533[Web of Science][Medline].
-
Takei N,
Endo Y
(1994)
Ca2+ ionophore-induced apoptosis on cultured embryonic rat cortical neurons.
Brain Res
652:65-70[Web of Science][Medline].
-
Thinakaran G,
Borchelt DR,
Lee MK,
Slunt HH,
Spitzer L,
Kim G,
Ratovitsky T,
Davenport F,
Norstedt C,
Seeger M,
Hardy J,
Levey A,
Gandy SE,
Jenkins NA,
Copeland NG,
Price DL,
Sisodia SA
(1996)
Endoproteolysis of presenilin-1 and accumulation of processed derivatives in vivo.
Neuron
17:181-190[Web of Science][Medline].
-
Thompson CB
(1995)
Apoptosis in the pathogenesis and treatment of disease.
Science
267:1456-1462[Abstract/Free Full Text].
-
Vito P,
Lacan E,
D'Adamio L
(1996)
Interfering with apoptosis: Ca2+-binding protein ALG-2 and Alzheimer's disease gene ALG-3.
Science
271:521-525[Abstract].
-
Walter J,
Capell A,
Grunberg J,
Pesold B,
Schindzielorz A,
Prior R,
Podlisny MB,
Fraser P,
St. George-Hyslop PH,
Selkoe DJ,
Haass C
(1996)
The Alzheimer's disease-associated presenilins are differentially phosphorylated proteins located predominantly within the endoplasmic reticulum.
Mol Med
2:673-691[Web of Science][Medline].
-
Weiss JH,
Pike CJ,
Cotman CW
(1994)
Ca2+ channel blockers attenuate -amyloid peptide toxicity to cortical neurons in culture.
J Neurochem
62:372-375[Web of Science][Medline].
-
Wolozin B,
Iwasaki K,
Vito P,
Ganjei JK,
Lacana E,
Sunderland T,
Zhao B,
Kusiak JW,
Wasco W,
D'Adamio L
(1996)
Participation of presenilin-2 in apoptosis: enhanced basal activity conferred by an Alzheimer mutation.
Science
274:1710-1713[Abstract/Free Full Text].
-
Zhang Y,
Tatsuno T,
Carney J,
Mattson MP
(1993)
Basic FGF, NGF, and IGFs protect hippocampal neurons against iron-induced degeneration.
J Cereb Blood Flow Metab
13:378-388[Web of Science][Medline].
This article has been cited by other articles:
|
|
|
|
 
B. Zhang, Y. Dong, G. Zhang, R. D. Moir, W. Xia, Y. Yue, M. Tian, D. J. Culley, G. Crosby, R. E. Tanzi, et al.
The Inhalation Anesthetic Desflurane Induces Caspase Activation and Increases Amyloid {beta}-Protein Levels under Hypoxic Conditions
J. Biol. Chem.,
May 2, 2008;
283(18):
11866 - 11875.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Liang, Q. Wang, Y. Li, B. Kang, M. F. Eckenhoff, R. G. Eckenhoff, and H. Wei
A Presenilin-1 Mutation Renders Neurons Vulnerable to Isoflurane Toxicity
Anesth. Analg.,
February 1, 2008;
106(2):
492 - 500.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Baki, R. L. Neve, Z. Shao, J. Shioi, A. Georgakopoulos, and N. K Robakis
Wild-Type But Not FAD Mutant Presenilin-1 Prevents Neuronal Degeneration by Promoting Phosphatidylinositol 3-Kinase Neuroprotective Signaling
J. Neurosci.,
January 9, 2008;
28(2):
483 - 490.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Xie, Y. Dong, U. Maeda, R. D. Moir, W. Xia, D. J. Culley, G. Crosby, and R. E. Tanzi
The Inhalation Anesthetic Isoflurane Induces a Vicious Cycle of Apoptosis and Amyloid {beta}-Protein Accumulation
J. Neurosci.,
February 7, 2007;
27(6):
1247 - 1254.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Xie and Q. Guo
Par-4 is a novel mediator of renal tubule cell death in models of ischemia-reperfusion injury
Am J Physiol Renal Physiol,
January 1, 2007;
292(1):
F107 - F115.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Estrada, A. Varshney, and B. E. Ehrlich
Elevated Testosterone Induces Apoptosis in Neuronal Cells
J. Biol. Chem.,
September 1, 2006;
281(35):
25492 - 25501.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Cai, P. Lin, K.-H. Cheung, N. Li, C. Levchook, Z. Pan, C. Ferrante, G. L. Boulianne, J. K. Foskett, D. Danielpour, et al.
The Presenilin-2 Loop Peptide Perturbs Intracellular Ca2+ Homeostasis and Accelerates Apoptosis
J. Biol. Chem.,
June 16, 2006;
281(24):
16649 - 16655.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
O. Lazarov, L. D. Peterson, D. A. Peterson, and S. S. Sisodia
Expression of a Familial Alzheimer's Disease-Linked Presenilin-1 Variant Enhances Perforant Pathway Lesion-Induced Neuronal Loss in the Entorhinal Cortex
J. Neurosci.,
January 11, 2006;
26(2):
429 - 434.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. P. Mattson and M. Gleichmann
The Neuronal Death Protein Par-4 Mediates Dopaminergic Synaptic Plasticity
Mol. Interv.,
October 1, 2005;
5(5):
278 - 281.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-L. Teo, H. Ma, C. Nguyen, C. Lam, and M. Kahn
Specific inhibition of CBP/{beta}-catenin interaction rescues defects in neuronal differentiation caused by a presenilin-1 mutation
PNAS,
August 23, 2005;
102(34):
12171 - 12176.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H.-Q. Wang, Y. Nakaya, Z. Du, T. Yamane, M. Shirane, T. Kudo, M. Takeda, K. Takebayashi, Y. Noda, K. I. Nakayama, et al.
Interaction of presenilins with FKBP38 promotes apoptosis by reducing mitochondrial Bcl-2
Hum. Mol. Genet.,
July 1, 2005;
14(13):
1889 - 1902.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. M. P. Ribeiro, A. M. Paradiso, U. Schwab, J. Perez-Vilar, L. Jones, W. O'Neal, and R. C. Boucher
Chronic Airway Infection/Inflammation Induces a Ca2+i-dependent Hyperinflammatory Response in Human Cystic Fibrosis Airway Epithelia
J. Biol. Chem.,
May 6, 2005;
280(18):
17798 - 17806.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Xie and Q. Guo
PAR-4 Is Involved in Regulation of {beta}-Secretase Cleavage of the Alzheimer Amyloid Precursor Protein
J. Biol. Chem.,
April 8, 2005;
280(14):
13824 - 13832.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. M. P. Ribeiro, A. M. Paradiso, M. A. Carew, S. B. Shears, and R. C. Boucher
Cystic Fibrosis Airway Epithelial Ca2+i Signaling: THE MECHANISM FOR THE LARGER AGONIST-MEDIATED Ca2+i SIGNALS IN HUMAN CYSTIC FIBROSIS AIRWAY EPITHELIA
J. Biol. Chem.,
March 18, 2005;
280(11):
10202 - 10209.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Verkhratsky
Physiology and Pathophysiology of the Calcium Store in the Endoplasmic Reticulum of Neurons
Physiol Rev,
January 1, 2005;
85(1):
201 - 279.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. A. Christensen, A. Shtifman, P. D. Allen, J. R. Lopez, and H. W. Querfurth
Calcium Dyshomeostasis in {beta}-Amyloid and Tau-bearing Skeletal Myotubes
J. Biol. Chem.,
December 17, 2004;
279(51):
53524 - 53532.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Verdile, A. Gnjec, J. Miklossy, J. Fonte, G. Veurink, K. Bates, B. Kakulas, P. D. Mehta, E. A. Milward, N. Tan, et al.
Protein markers for Alzheimer disease in the frontal cortex and cerebellum
Neurology,
October 26, 2004;
63(8):
1385 - 1392.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. L. Chan, W. Fu, P. Zhang, A. Cheng, J. Lee, K. Kokame, and M. P. Mattson
Herp Stabilizes Neuronal Ca2+ Homeostasis and Mitochondrial Function during Endoplasmic Reticulum Stress
J. Biol. Chem.,
July 2, 2004;
279(27):
28733 - 28743.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Hashimoto, O. Tsuji, K. Kanekura, S. Aiso, T. Niikura, M. Matsuoka, and I. Nishimoto
The Gtx Homeodomain Transcription Factor Exerts Neuroprotection Using Its Homeodomain
J. Biol. Chem.,
April 16, 2004;
279(16):
16767 - 16777.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Vaghefi, A. L. Hughes, and K. E. Neet
Nerve Growth Factor Withdrawal-mediated Apoptosis in Naive and Differentiated PC12 Cells through p53/Caspase-3-dependent and -independent Pathways
J. Biol. Chem.,
April 9, 2004;
279(15):
15604 - 15614.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. A. Marques, U. Keil, A. Bonert, B. Steiner, C. Haass, W. E. Muller, and A. Eckert
Neurotoxic Mechanisms Caused by the Alzheimer's Disease-linked Swedish Amyloid Precursor Protein Mutation: OXIDATIVE STRESS, CASPASES, AND THE JNK PATHWAY
J. Biol. Chem.,
July 18, 2003;
278(30):
28294 - 28302.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Kitagawa, S. Shimohama, T. Oeda, K. Uemura, R. Kohno, A. Kuzuya, H. Shibasaki, and N. Ishii
The Role of the Presenilin-1 Homologue Gene sel-12 of Caenorhabditis elegans in Apoptotic Activities
J. Biol. Chem.,
March 28, 2003;
278(14):
12130 - 12134.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Fath, J. Eidenmuller, and R. Brandt
Tau-Mediated Cytotoxicity in a Pseudohyperphosphorylation Model of Alzheimer's Disease
J. Neurosci.,
November 15, 2002;
22(22):
9733 - 9741.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Song, P. De Sarno, and R. S. Jope
Central Role of Glycogen Synthase Kinase-3beta in Endoplasmic Reticulum Stress-induced Caspase-3 Activation
J. Biol. Chem.,
November 15, 2002;
277(47):
44701 - 44708.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Cedazo-Minguez, B. O. Popescu, M. Ankarcrona, T. Nishimura, and R. F. Cowburn
The Presenilin 1 Delta E9 Mutation Gives Enhanced Basal Phospholipase C Activity and a Resultant Increase in Intracellular Calcium Concentrations
J. Biol. Chem.,
September 20, 2002;
277(39):
36646 - 36655.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y.-H. Suh and F. Checler
Amyloid Precursor Protein, Presenilins, and alpha -Synuclein: Molecular Pathogenesis and Pharmacological Applications in Alzheimer's Disease
Pharmacol. Rev.,
September 1, 2002;
54(3):
469 - 525.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. P. Mattson, S. L. Chan, and W. Duan
Modification of Brain Aging and Neurodegenerative Disorders by Genes, Diet, and Behavior
Physiol Rev,
July 1, 2002;
82(3):
637 - 672.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Lu, S. L. Chan, W. Fu, and M. P. Mattson
The Lipid Peroxidation Product 4-Hydroxynonenal Facilitates Opening of Voltage-dependent Ca2+ Channels in Neurons by Increasing Protein Tyrosine Phosphorylation
J. Biol. Chem.,
June 28, 2002;
277(27):
24368 - 24375.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. E. Tarr, R. Roncarati, G. Pelicci, P. G. Pelicci, and L. D'Adamio
Tyrosine Phosphorylation of the beta -Amyloid Precursor Protein Cytoplasmic Tail Promotes Interaction with Shc
J. Biol. Chem.,
May 3, 2002;
277(19):
16798 - 16804.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Perkins, E. F. R. Pereira, M. Gober, P. J. Yarowsky, and L. Aurelian
The Herpes Simplex Virus Type 2 R1 Protein Kinase (ICP10 PK) Blocks Apoptosis in Hippocampal Neurons, Involving Activation of the MEK/MAPK Survival Pathway
J. Virol.,
February 1, 2002;
76(3):
1435 - 1449.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Katayama, K. Imaizumi, A. Honda, T. Yoneda, T. Kudo, M. Takeda, K. Mori, R. Rozmahel, P. Fraser, P. St. George-Hyslop, et al.
Disturbed Activation of Endoplasmic Reticulum Stress Transducers by Familial Alzheimer's Disease-linked Presenilin-1 Mutations
J. Biol. Chem.,
November 9, 2001;
276(46):
43446 - 43454.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X.-L. Ding, J. Husseman, A. Tomashevski, D. Nochlin, L.-W. Jin, and I. Vincent
The Cell Cycle Cdc25A Tyrosine Phosphatase Is Activated in Degenerating Postmitotic Neurons in Alzheimer's Disease
Am. J. Pathol.,
December 1, 2000;
157(6):
1983 - 1990.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. L. Mah, G. Perry, M. A. Smith, and M. J. Monteiro
Identification of Ubiquilin, a Novel Presenilin Interactor That Increases Presenilin Protein Accumulation
J. Cell Biol.,
November 13, 2000;
151(4):
847 - 862.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Grilli, E. Diodato, G. Lozza, R. Brusa, M. Casarini, D. Uberti, R. Rozmahel, D. Westaway, P. St George-Hyslop, M. Memo, et al.
Presenilin-1 regulates the neuronal threshold to excitotoxicity both physiologically and pathologically
PNAS,
November 7, 2000;
97(23):
12822 - 12827.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Leissring, T. R. Yamasaki, W. Wasco, J. D. Buxbaum, I. Parker, and F. M. LaFerla
Calsenilin reverses presenilin-mediated enhancement of calcium signaling
PNAS,
July 18, 2000;
97(15):
8590 - 8593.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. M. Cataldo, C. M. Peterhoff, J. C. Troncoso, T. Gomez-Isla, B. T. Hyman, and R. A. Nixon
Endocytic Pathway Abnormalities Precede Amyloid {beta} Deposition in Sporadic Alzheimer's Disease and Down Syndrome : Differential Effects of APOE Genotype and Presenilin Mutations
Am. J. Pathol.,
July 1, 2000;
157(1):
277 - 286.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Takeuchi, M. C. Irizarry, K. Duff, T. C. Saido, K. Hsiao Ashe, M. Hasegawa, D. M. A. Mann, B. T. Hyman, and T. Iwatsubo
Age-Related Amyloid {beta} Deposition in Transgenic Mice Overexpressing Both Alzheimer Mutant Presenilin 1 and Amyloid {beta} Precursor Protein Swedish Mutant Is Not Associated with Global Neuronal Loss
Am. J. Pathol.,
July 1, 2000;
157(1):
331 - 339.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Amson, J.-M. Lassalle, H. Halley, S. Prieur, F. Lethrosne, J.-P. Roperch, D. Israeli, M.-C. Gendron, C. Duyckaerts, F. Checler, et al.
Behavioral alterations associated with apoptosis and down-regulation of presenilin 1 in the brains of p53-deficient mice
PNAS,
May 9, 2000;
97(10):
5346 - 5350.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A B Harkins, S Dlouhy, B Ghetti, A L Cahill, L Won, B Heller, A Heller, and A P Fox
Evidence of elevated intracellular calcium levels in weaver homozygote mice
J. Physiol.,
April 15, 2000;
524(2):
447 - 455.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. P. Mattson, H. Zhu, J. Yu, and M. S. Kindy
Presenilin-1 Mutation Increases Neuronal Vulnerability to Focal Ischemia In Vivo and to Hypoxia and Glucose Deprivation in Cell Culture: Involvement of Perturbed Calcium Homeostasis
J. Neurosci.,
February 15, 2000;
20(4):
1358 - 1364.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. J. Palacino, B. E. Berechid, P. Alexander, C. Eckman, S. Younkin, J. S. Nye, and B. Wolozin
Regulation of Amyloid Precursor Protein Processing by Presenilin 1 (PS1) and PS2 in PS1 Knockout Cells
J. Biol. Chem.,
January 7, 2000;
275(1):
215 - 222.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Leissring, I. Parker, and F. M. LaFerla
Presenilin-2 Mutations Modulate Amplitude and Kinetics of Inositol 1,4,5-Trisphosphate-mediated Calcium Signals
J. Biol. Chem.,
November 12, 1999;
274(46):
32535 - 32538.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X. Xu, Y.-c. Shi, X. Wu, P. Gambetti, D. Sui, and M.-Z. Cui
Identification of a Novel PSD-95/Dlg/ZO-1 (PDZ)-like Protein Interacting with the C Terminus of Presenilin-1
J. Biol. Chem.,
November 12, 1999;
274(46):
32543 - 32546.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Stadelmann, T. L. Deckwerth, A. Srinivasan, C. Bancher, W. Bruck, K. Jellinger, and H. Lassmann
Activation of Caspase-3 in Single Neurons and Autophagic Granules of Granulovacuolar Degeneration in Alzheimer’s Disease : Evidence for Apoptotic Cell Death
Am. J. Pathol.,
November 1, 1999;
155(5):
1459 - 1466.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Herreman, D. Hartmann, W. Annaert, P. Saftig, K. Craessaerts, L. Serneels, L. Umans, V. Schrijvers, F. Checler, H. Vanderstichele, et al.
Presenilin 2 deficiency causes a mild pulmonary phenotype and no changes in amyloid precursor protein processing but enhances the embryonic lethal phenotype of presenilin 1 deficiency
PNAS,
October 12, 1999;
96(21):
11872 - 11877.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Chierzi, E. Strettoi, M. C. Cenni, and L. Maffei
Optic Nerve Crush: Axonal Responses in Wild-Type and bcl-2 Transgenic Mice
J. Neurosci.,
October 1, 1999;
19(19):
8367 - 8376.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Ye and M. E. Fortini
Apoptotic Activities of Wild-Type and Alzheimer's Disease-Related Mutant Presenilins in Drosophila melanogaster
J. Cell Biol.,
September 20, 1999;
146(6):
1351 - 1364.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Gomez-Isla, W. B. Growdon, M. J. McNamara, D. Nochlin, T. D. Bird, J. C. Arango, F. Lopera, K. S. Kosik, P. L. Lantos, N. J. Cairns, et al.
The impact of different presenilin 1 andpresenilin 2 mutations on amyloid deposition, neurofibrillary changes and neuronal loss in the familial Alzheimer's disease brain: Evidence for other phenotype-modifying factors
Brain,
September 1, 1999;
122(9):
1709 - 1719.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. J. Passer, L. Pellegrini, P. Vito, J. K. Ganjei, and L. D'Adamio
Interaction of Alzheimer's Presenilin-1 and Presenilin-2 with Bcl-XL. A POTENTIAL ROLE IN MODULATING THE THRESHOLD OF CELL DEATH
J. Biol. Chem.,
August 20, 1999;
274(34):
24007 - 24013.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Pastorcic and H. K. Das
An Upstream Element Containing an ETS Binding Site Is Crucial for Transcription of the Human Presenilin-1 Gene
J. Biol. Chem.,
August 20, 1999;
274(34):
24297 - 24307.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Pellegrini, B. J. Passer, M. Tabaton, J. K. Ganjei, and L. D'Adamio
Alternative, Non-secretase Processing of Alzheimer's beta -Amyloid Precursor Protein during Apoptosis by Caspase-6 and -8
J. Biol. Chem.,
July 23, 1999;
274(30):
21011 - 21016.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. L. Pahl
Signal Transduction From the Endoplasmic Reticulum to the Cell Nucleus
Physiol Rev,
July 1, 1999;
79(3):
683 - 701.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. M. Stabler, L. L. Ostrowski, S. M. Janicki, and M. J. Monteiro
A Myristoylated Calcium-binding Protein that Preferentially Interacts with the Alzheimer's Disease Presenilin 2 Protein
J. Cell Biol.,
June 14, 1999;
145(6):
1277 - 1292.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. J. Kaufman
Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls
Genes & Dev.,
May 15, 1999;
13(10):
1211 - 1233.
[Full Text]
|
|
|
|
|
|
|
Q. Guo, L. Sebastian, B. L. Sopher, M. W. Miller, G. W. Glazner, C. B. Ware, G. M. Martin, and M. P. Mattson
Neurotrophic factors [activity-dependent neurotrophic factor (ADNF) and basic fibroblast growth factor (bFGF)] interrupt excitotoxic neurodegenerative cascades promoted by a PS1 mutation
PNAS,
March 30, 1999;
96(7):
4125 - 4130.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Imaizumi, T. Morihara, Y. Mori, T. Katayama, M. Tsuda, T. Furuyama, A. Wanaka, M. Takeda, and M. Tohyama
The Cell Death-promoting Gene DP5, Which Interacts with the BCL2 Family, Is Induced during Neuronal Apoptosis Following Exposure to Amyloid beta Protein
J. Biol. Chem.,
March 19, 1999;
274(12):
7975 - 7981.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. Fu, J. G. Begley, M. W. Killen, and M. P. Mattson
Anti-apoptotic Role of Telomerase in Pheochromocytoma Cells
J. Biol. Chem.,
March 12, 1999;
274(11):
7264 - 7271.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. P. Mattson and Qing Guo
{blacksquare} REVIEW : The Presenilins
Neuroscientist,
March 1, 1999;
5(2):
112 - 124.
[Abstract]
[PDF]
|
|
|
|
|
|
|
C. Choi, J. Y. Park, J. Lee, J.-H. Lim, E.-C. Shin, Y. S. Ahn, C.-H. Kim, S.-J. Kim, J.-D. Kim, I. S. Choi, et al.
Fas Ligand and Fas Are Expressed Constitutively in Human Astrocytes and the Expression Increases with IL-1, IL-6, TNF-{alpha}, or IFN-{gamma}
J. Immunol.,
February 15, 1999;
162(4):
1889 - 1895.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C.-S. Hong, L. Caromile, Y. Nomata, H. Mori, D. E. Bredesen, and E. H. Koo
Contrasting Role of Presenilin-1 and Presenilin-2 in Neuronal Differentiation In Vitro
J. Neurosci.,
January 15, 1999;
19(2):
637 - 643.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Jaikaran, G Marcon, L Levesque, P. George-Hyslop, P. Fraser, and A Clark
Localisation of presenilin 2 in human and rodent pancreatic islet beta-cells; Met239Val presenilin 2 variant is not associated with diabetes in man
J. Cell Sci.,
January 7, 1999;
112(13):
2137 - 2144.
[Abstract]
[PDF]
|
|
|
|
|
|
|
S. Bursztajn, R. DeSouza, D. L. McPhie, S. A. Berman, J. Shioi, N. K. Robakis, and R. L. Neve
Overexpression in Neurons of Human Presenilin-1 or a Presenilin-1 Familial Alzheimer Disease Mutant Does Not Enhance Apoptosis
J. Neurosci.,
December 1, 1998;
18(23):
9790 - 9799.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Takashima, M. Murayama, O. Murayama, T. Kohno, T. Honda, K. Yasutake, N. Nihonmatsu, M. Mercken, H. Yamaguchi, S. Sugihara, et al.
Presenilin 1 associates with glycogen synthase kinase-3beta and its substrate tau
PNAS,
August 4, 1998;
95(16):
9637 - 9641.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. N. Keller, Q. Guo, F. W. Holtsberg, A. J. Bruce-Keller, and M. P. Mattson
Increased Sensitivity to Mitochondrial Toxin-Induced Apoptosis in Neural Cells Expressing Mutant Presenilin-1 Is Linked to Perturbed Calcium Homeostasis and Enhanced Oxyradical Production
J. Neurosci.,
June 15, 1998;
18(12):
4439 - 4450.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Guo, N. Robinson, and M. P. Mattson
Secreted beta -Amyloid Precursor Protein Counteracts the Proapoptotic Action of Mutant Presenilin-1 by Activation of NF-kappa B and Stabilization of Calcium Homeostasis
J. Biol. Chem.,
May 15, 1998;
273(20):
12341 - 12351.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Guo, S. Christakos, N. Robinson, and M. P. Mattson
Calbindin D28k blocks the proapoptotic actions of mutant presenilin 1: Reduced oxidative stress and preserved mitochondrial function
PNAS,
March 17, 1998;
95(6):
3227 - 3232.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. A. Pedersen, Q. Guo, B. K. Hartman, and M. P. Mattson
Nerve Growth Factor-independent Reduction in Choline Acetyltransferase Activity in PC12 Cells Expressing Mutant Presenilin-1
J. Biol. Chem.,
September 5, 1997;
272(36):
22397 - 22400.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. L. Chan, M. Mayne, C. P. Holden, J. D. Geiger, and M. P. Mattson
Presenilin-1 Mutations Increase Levels of Ryanodine Receptors and Calcium Release in PC12 Cells and Cortical Neurons
J. Biol. Chem.,
June 9, 2000;
275(24):
18195 - 18200.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Schneider, D. Reverse, I. Dewachter, L. Ris, N. Caluwaerts, C. Kuiperi, M. Gilis, H. Geerts, H. Kretzschmar, E. Godaux, et al.
Mutant Presenilins Disturb Neuronal Calcium Homeostasis in the Brain of Transgenic Mice, Decreasing the Threshold for Excitotoxicity and Facilitating Long-term Potentiation
J. Biol. Chem.,
April 6, 2001;
276(15):
11539 - 11544.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Guo, J. Xie, X. Chang, and H. Du
Prostate Apoptosis Response-4 Enhances Secretion of Amyloid beta Peptide 1-42 in Human Neuroblastoma IMR-32 Cells by a Caspase-dependent Pathway
J. Biol. Chem.,
May 4, 2001;
276(19):
16040 - 16044.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Pan, M. B. Bhat, A.-L. Nieminen, and J. Ma
Synergistic Movements of Ca2+ and Bax in Cells Undergoing Apoptosis
J. Biol. Chem.,
August 17, 2001;
276(34):
32257 - 32263.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. H. Scheinfeld, R. Roncarati, P. Vito, P. A. Lopez, M. Abdallah, and L. D'Adamio
Jun NH2-terminal Kinase (JNK) Interacting Protein 1 (JIP1) Binds the Cytoplasmic Domain of the Alzheimer's beta -Amyloid Precursor Protein (APP)
J. Biol. Chem.,
January 25, 2002;
277(5):
3767 - 3775.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Pan, D. Damron, A.-L. Nieminen, M. B. Bhat, and J. Ma
Depletion of Intracellular Ca2+ by Caffeine and Ryanodine Induces Apoptosis of Chinese Hamster Ovary Cells Transfected with Ryanodine Receptor
J. Biol. Chem.,
June 23, 2000;
275(26):
19978 - 19984.
[Abstract]
[Full Text]
[PDF]
|
|
|
Compound via MeSH
