neuroprotective effects of memantine
DESCRIPTION
Neuroprotective Effects of Memantine. Parallel incubation of 3-NP and memantine for 7 or 12 days. Cresyl violet staining. Homogenization Immunoblot Incubation with synaptic markers. Memantine in an In Vitro Model for Neurodegeneration. - PowerPoint PPT PresentationTRANSCRIPT
Neuroprotective Effects of
Memantine
Hippocampal slice cultures
Brown et al., Soc. Neurosci 2003
Semi-chronic 3-NP toxicity in organotypic hippocampal cultures
Parallel incubation of 3-NP and memantine for 7 or 12 days
Cresyl violet staining
Homogenization Immunoblot Incubation with
synaptic markers
Memantine in an In Vitro Model for Neurodegeneration
Semi-chronic 3-NP toxicity in organotypic hippocampal cultures
Brown et al., Soc. Neurosci 2003
3-NP for 7 days300
200
100
0Control 10 µM1 µM 5 µM
MemantineVehicle
Glu
R1
(a
rbit
ary
un
its
)Neuroprotective Effect of Memantine
In Vitro
Semi-chronic 3-NP toxicity in organotypic hippocampal cultures
Brown et al., Soc. Neurosci 2003
3-NP for 7 days300
200
100
0VehicleControl 10 µM1 µM 5 µM
Memantine
Sy
na
ps
in II
b (
arb
itar
y u
nit
s)
Neuroprotective Effect of MemantineIn Vitro
CA1
Control 3-NP 14 days 3-NP + Memantine
DG
Semi-chronic 3-NP toxicity in organotypic hippocampal cultures
Brown et al., Soc. Neurosci 2003
Neuroprotective Effect of MemantineIn Vitro
orMemantine injection 30 min before NMDA
Memantine infusion for 2 weeks
Unilateral injection of NMDA (7.5 nmol) or 3NP (250 nmol) into the NBM
Biochemical assay
Choline acetyltransferase in the frontal cortex
2 weeks
Wenk et al., Eur J Pharmacol 1995, NeuroReport 1996
Memantine‘s effect on lesions of the nucleus basalis magnocellularis (NBM)
Memantine in an Animal Model for Neurodegenerative Dementia
Ch
AT
act
ivit
y c
on
tro
l - le
sio
ne
d s
ide
(n
mo
l AC
h/h
*mg
pro
tein
e)
Wenk et al., Eur J Pharmacol 1995
8
6
4
2
0
10.01 0.1 10 100 1000
Memantine (ED50 = 2.8 mg/kg)
MK-801 (ED50 = 0.077 mg/kg)
Dose (mg/kg)
Memantine injection (i.p.) attenuated NMDA-induced lesion of the NBM
Protection of Cholinergic Neurons by Memantine
Wenk et al., NeuroReport 1996
Co
rtic
al C
hA
T a
cti
vit
y c
on
tro
l - le
sio
ne
d s
ide
(n
mo
l AC
h/h
*mg
pro
tein
e)
* p < 0.01 versus control
3-NPlesion
NMDAlesion
0
-4
-8
-12
-16
-20
*
* Memantine
(20 mg/kg/d)
Control
Infusion of memantine attenuated damage to NBM neurones induced by injection of NMDA or 3-NP
Degeneration of Cholinergic Neurons was Attenuated by Memantine
Memantine infusion(20 mg/kg/day)
7 days Immunohistochemistry in the hippocampus:
neuronal damage
GFAP
Miguel-Hidalgo et al., Brain Res 2002
Injection of ß-amyloid (1-40)into the hippocampus
2 days
Memantine‘s effect on ß-amyloid-induced lesion of the hippocampus
Memantine in an Animal Model for Alzheimer´s Disease
Memantine
Placebo Extent of ß-amyloid-induced damage in the CA1 region
1000
800
600
400
200
0
Ex
ten
t (µ
m)
Vehicle Memantine
*
Miguel-Hidalgo et al., Brain Res 2002
* p < 0.02 versus placebo
Protection by Memantine Against A-Induced Neurodegeneration
Vehicle
Memantine
Miguel-Hidalgo et al., Brain Res 2002
* p < 0.03 versus vehicle
Are
a (
%)
Protection by Memantine Against A-Induced Neurodegeneration
Area of GFAP profilesaround the injection site
30
25
20
15
10
5
0
Vehicle Memantine
*
Vehicle Memantine
Memantine effect on lipopolysaccharide (LPS)-induced brain insult and inflammation
Memantine infusion(s.c. 20 mg/kg/day for 37 days)
Infusion of LPSinto the basal forebrain (37 days)
Willard et al., Exp Brain Res 2000
Biochemical analysis in the frontal cortex:
ChAT activity
10 days
Effect of Memantine on Inflammation Induced Neurodegeneration
* p < 0.0001 versus control; ** p < 0.05 versus LPS
Willard et al., Exp Brain Res 2000
Effect of LPS infusion into the basal forebrain on cortical ChAT activity
5
0
-5
-10
-15
-20
-25
**
Control LPS LPS + Memantine
*De
clin
e in
co
rtic
al C
hA
T
act
ivit
y [%
]Memantine Protected Cholinergic
Neurons from Damage by Inflammation
Effects of memantine on quinolinic acid-induced neurodegeneration
15 sec
10 trials/day5 days
T-maze alternation
Biochemicalanalysis
Removal of minipumps
3 days
Parallel infusion of memantine (s.c. 20 mg/kg/day) and
quinolinic acid (i.c.v.)
2 weeks
Misztal et al., Eur J Pharmacol 1996
Memantine in an Animal Model for Neurodegenerative Dementia
Infusion of Memantine (20 mg/kg/day) attenuated T-maze learning deficit induced by chronic i.c.v. infusion of quinolinic acid (QA)
Day
Misztal et al., Eur J Pharmacol 1996
* p < 0.05 versus control and QA + Memantine
Control QA QA + Memantine
4
3
2
1
01 532 4
*
* **
*
Nu
mb
er
of
erro
rsAttenuation of Quinolinic Acid Induced
Memory Loss by Memantine
Memantine (20 mg/kg/day) attenuated the hippocampal cholinergic deficit induced by chronic i.c.v. infusion of quinolinic acid (QA)
* p < 0.05 versus quinolinic acid
**
100
80
60
40
20
0Control QA QA + Memantine
Misztal et al., Eur J Pharmacol 1996
[H3]H
em
ich
olin
ium
-3
bin
din
g (
µm
ol/m
g t
iss
ue)
Attenuation of Quinolinic Acid Induced Neurodegeneration by Memantine
Summary
Neuroprotective effects of memantine were shown,
in vivo on
Excitotoxic induced neurodegeneration
ß-amyloid induced neuronal damage
LPS induced inflammation
in vitro on
Metabolic disturbances due to mitochondrial dysfunction
Conclusion:
Neuroprotective effects of memantine have been shown under
various conditions which are clinically relevant for Alzheimer's
disease.
Memantine Inhibits and Reverses the
Alzheimer Type Abnormal
Hyperphosphorylation of tau and
Associated Neurodegeneration
Li L., Sengupta A., Haque N., Grundke-Iqbal I. and Iqbal K.
FEBS Letters, 2004, 566 (1-3): 261-269
Tau Hyperphosphorylation in Alzheimer‘s Disease
Alzheimer‘s disease In vitro model
Hippocampal culture
+ okadaic acid (OA)
PP-2A activity
CaMKII activity
PKA activity
Hyperphosphorylation
of tau
Therapeutic approach
Hyperphosphorylation
of tau
Tangle formation
Neurodegeneration
Okadaic acid for 24h Memantine or vehicle for 24h
Hippocampal slices
Analysis
Effects of Memantine on Phosphorylation of tau - Methods
Assay for phosphatase- or kinase activity
Western blots (p-tau)
Assay for cell death
Memantine Counteracted OA-induced PP-2A Inhibition
100 100 100 0 0
PP-2A activity
0 1 10 1 10
PP
-2A
ac
tivi
ty (
% o
f c
on
tro
l)
24 h OA + 24 h vehicle 24 h OA + 24 h Mem 24 h Mem
* p < 0.05 versus OA treated tissue
120
100
80
60
40
20
0nM OA
µM Mem
*
CaMKII activity PKA activity
Memantine Restored CaMKII and PKA Activity
Kin
ase
ac
tiv
ity
( %
of
co
ntr
ol)
* p < 0.05 versus OA treated tissue
250
200
150
100
50
0
100 100 100 0 0
120
100
80
60
40
20
0
0 1 10 1 10
nM OA µM Mem
100 100 100 0 0 0 1 10 1 10
24 h OA + 24 h vehicle 24 h OA + 24 h Mem 24 h Mem
*
**
Memantine Counteracted OA-induced Cell Death
10
8
6
4
2
0
Cell death assay
Control 24 h OA + 24 h vehicle 24 h OA + 24 h Mem 24 h Mem
LD
H r
ele
as
e(r
ati
o a
fte
r / b
efo
re t
rea
tme
nt)
nM OA 0 100 100 100 0 0
µM Mem 0 0 1 10 1 10
* *
* p < 0.05 versus OA treated tissue
3 21
68
43
100 nM OA - + + 10 µM Mem - - +
Memantine Counteracted CaMKII-induced Phosphorylation of tau
[125I] Western blot with Antibody against pS-262
Control 24 h OA + 24 h vehicle 24 h OA + 24 h Mem (10 µM)
Ph
osp
ho
ryla
tio
n o
f ta
u
7
6
5
4
3
2
1
pSer262 pSer422
Immunocytochemical staining of pSer-262
100 nM OA + + + 10 µM Mem - - -
100 nM OA - + + 10 µM Mem - - +
Memantine Counteracted OA-induced Phosphorylation of tau
In an in vitro model using okadaic acid memantine was
shown to
Restore normal PP-2A, CaMKII and PKA activities
Prevent cell death
Positively influence phosphorylation / dephosphorylation
imbalance
Conclusion
Apart from the symptomatic benefits, memantine might also
positively influence pathological changes in AD.
Summary