Effects of physical exercise in ageing and Alzheimer’s disease markers

1.

Summary results
IIBB-CSIC
Febrer 2016

2. 5xFAD + Physical Exercise

2 month-old male or
female mice
Males (n)
Females (n)
WT (12)
WT (7)
5xFAD Sedentary (6)
5xFAD Sedentary (7)
5xFAD Exercise (7)
5xFAD Exercise (7)
6 months of treatment
Behavioral tests (BPSDs and cognitive behavior)
IHC and biochemical tests (WB and PCR)

3. 5xFAD + Physical Exercise

Behavioral tests:
Open field
Dark and light box test
neophobia
anxiety
4-hole board test
Latència exploració 4 forats
*
*
**
30
Temps (s)
exploratory
capacity
Latència (s)
300
Temps exploracions
200
100
**
20
10
0
0
WT
5xFAD Sed
*p<0.05; **p<0.01; ****p<0.0001
5xFAD Exe
WT
5xFAD Sed
5xFAD Exe

4. 5xFAD + Physical Exercise

Behavioral tests:
NORT 0h - Índex de discriminació
0.5
0.4
0.3
0.2
0.1
0.0
-0.1
WT
5xFAD Sed
5xFAD Exe
Índex de discriminació
0.5
Índex de discriminació
Índex de discriminació
0.5
-0.2
NORT 24h - Índex de discriminació
NORT 2h - Índex de discriminació
0.4
0.3
0.2
0.1
0.0
0.4
0.3
0.2
0.1
0.0
WT
5xFAD Sed
5xFAD Exe
Physical exercise restores cognitive
decline observed in 5xFAD mice
WT
5xFAD Sed
5xFAD Exe

5. 5xFAD + Physical Exercise

Immunohistochemistry:
Physical exercise
does not decrease
amyloid burden nor
glial activation

6. 5xFAD + Physical Exercise

WB:
from mouse cortex
S y n a p to p h y s in - F e m a le s
P S D - 9 5 - F e m a le s
1 .5
1 .5
#
#
1 .0
p = 0 .0 5 5
N M D A R 2 B - F e m a le s
O .D .
O .D .
1 .0
1 .5
0 .5
0 .5
0 .0
0 .0
WT
5xFAD
WT
5xFAD E xe
O n e - w a y A N O V A : p = 0 .0 2 2 6
5xFAD
5xFAD E xe
O n e - w a y A N O V A : p = 0 .0 4 7 6
n = 6 -7 -6
n = 6 - 6 - 4 ( a u g m e n ta r n 5 x F A D E x e )
# p < 0 .0 5 v s . 5 x F A D
# p < 0 .0 5 v s . 5 x F A D
O .D .
1 .0
0 .5
0 .0
WT
P S D -9 5 - M a le s
S y n a p t o p h y s i n - M a le s
1 .5
1 .0
1 .0
O .D .
O .D .
1 .5
0 .5
0 .5
0 .0
0 .0
WT
5xFAD
5xFAD E xe
WT
5xFAD
5xFAD E xe
5xFAD
5xFAD E xe
C a l a u g m e n ta r n e n W T i 5 x F A D ( n = 4 - 4 - 5 )

7. 5xFAD + Physical Exercise

WB:
from mouse cortex
A D A M 1 0 m a tu r e
ADAM 10 FL
2 .0
S ir t - 1
1 .5
1 .5
1 .0
1 .0
O .D .
O .D .
1 .0
0 .5
0 .5
0 .5
0 .0
0 .0
WT
5xFAD
5xFAD E xe
0 .0
WT
n = 4 - 5 - 7 ( a u g m e n ta r n W T )
5xFAD
5xFAD E xe
WT
Ib a 1
2 .5
1 .5
**
*
2 .0
#
1 .0
O .D .
1 .5
1 .0
0 .5
0 .5
0 .0
0 .0
WT
5xFAD
5xFAD E xe
O n e - w a y A N O V A : p = 0 .0 0 9 4
n = 7 -8 -6
* * p < 0 .0 1 v s . W T
5xFAD
5xFAD E xe
n = 4 - 4 - 5 ( a u g m e n ta r n W T i 5 x F A D )
n = 4 -5 -6
G FAP
O .D .
O .D .
1 .5
WT
5xFAD
5xFAD E xe
O n e - w a y A N O V A : p = 0 .0 2 0 4
n = 3 - 4 - 6 ( a u g m e n ta r n W T i 5 x F A D )
* p < 0 .0 5 v s . W T
# p < 0 .0 5 v s . 5 x F A D

8. 5xFAD + Physical Exercise

PCR: from mouse hippocampus
TNF-a
****
****
****
Relative
expression
6
Testing results reproducibility…
Wt
Tg5XFAD
4
2
T N F - M a le s
s
th
on
m
T N F - F e m a le s (S Y B R )
T N F - F e m a le s
4
2 .0
*
3
*
2
1
0
R e la tiv e e x p r e s s io n
2 .0
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
8
2
m
on
th
s
0
1 .5
1 .0
0 .5
5xF AD
5xF AD E xe
1 .5
1 .0
0 .5
0 .0
0 .0
WT
*
WT
5xF AD
5xF AD E xe
WT
5xF AD
5xF AD E xe
O n e - w a y A N O V A : p = 0 .0 1 5 5
* p < 0 .0 5 v s . W T
1 5 u g c D N A /w e ll
C a rr e g a n t 1 5 u g c D N A p e r p o u
C a rr e g a n t 1 8 u g c D N A p e r p o u

9. 5xFAD + Physical Exercise

PCR: from mouse hippocampus
Testing RNA integrity…

10. 5xFAD + Physical Exercise

PCR: from mouse hippocampus (n=5-7)
B D N F - M a le s
N tr k 2 - M a le s
1 .5
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
1 .5
1 .0
0 .5
0 .0
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
WT
B D N F - F e m a le s
5xF AD E xe
N tr k 2 - F e m a le s
1 .5
R e la tiv e e x p r e s s io n
1 .5
R e la tiv e e x p r e s s io n
5xF AD
1 .0
0 .5
0 .0
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
WT
5xF AD
5xF AD E xe

11. 5xFAD + Physical Exercise

PCR: from mouse hippocampus (n=5-7)
S irt-1 - M a le s
A D A M 1 0 - M a le s
1 .5
1 .0
0 .5
2 .0
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
1 .5
1 .0
0 .5
0 .0
0 .0
WT
5xF AD
5xF AD
5xF AD E xe
WT
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
5xF AD
5xF AD E xe
5xF AD E xe
2 .0
1 .0
0 .5
0 .0
WT
5xF AD
G D N F - F e m a le s
1 .5
0 .0
0 .5
S ir t-1 - F e m a le s
1 .5
0 .5
1 .0
C a l re p e tir p e rq u è v a n s o r tir m o lts d u p lic a ts e s tra n y s
A D A M 1 0 - F e m a le s
1 .0
1 .5
0 .0
WT
5xF AD E xe
C a r re g a n t 3 .7 5 u g c D N A p e r p o u
R e la tiv e e x p r e s s io n
G D N F - M a le s
1 .5
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
WT
5xF AD
5xF AD E xe
C a l re p e tir p e rq u è v a n s o r tir m o lts d u p lic a ts e s tra n y s

12. 5xFAD + Physical Exercise

PCR: from mouse hippocampus (n=5-7)
H D A C 1 - M a le s
H D A C 2 - M a le s
1 .0
0 .5
0 .0
1 .5
R e la tiv e e x p r e s s io n
1 .5
R e la tiv e e x p r e s s io n
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
H D A C 1 - F e m a le s
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
5xF AD
5xF AD E xe
WT
0 .0
5xF AD
5xF AD E xe
5xF AD E xe
1 .5
1 .0
*
0 .5
1 .0
0 .5
0 .0
0 .0
WT
5xF AD
H D A C 5 - F e m a le s
1 .5
0 .5
0 .5
H D A C 2 - F e m a le s
1 .5
1 .0
1 .0
0 .0
WT
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
1 .5
H D A C 5 - M a le s
WT
5xF AD
5xF AD E xe
O n e - w a y A N O V A : p = 0 .0 4 1 2
* p < 0 .0 5 v s . W T
WT
5xF AD
5xF AD E xe

13. 5xFAD + Physical Exercise

PCR: from mouse hippocampus (n=5-7)
D n m t1 - M a le s
D n m t3 a - M a le s
1 .0
0 .5
0 .0
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
5xF AD
5xF AD E xe
WT
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
5xF AD E xe
1 .5
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
0 .0
5xF AD
D n m t3 b - F e m a le s
1 .5
0 .5
0 .5
D n m t3 a - F e m a le s
1 .5
1 .0
1 .0
0 .0
WT
D n m t1 - F e m a le s
R e la tiv e e x p r e s s io n
1 .5
R e la tiv e e x p r e s s io n
1 .5
R e la tiv e e x p r e s s io n
R e la tiv e e x p r e s s io n
1 .5
D n m t3 b - M a le s
1 .0
0 .5
0 .0
WT
5xF AD
5xF AD E xe
WT
5xF AD
5xF AD E xe