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“Ligandos Wnt como neurotrofinas, plasticidad sináptica modulada por ellos”
Dr. Jorge ParodiUniversidad Católica Temuco
Bubnik 2007 TRENDS
Schulte 2010, Phar- Review
Janda et al 2012 Science
Nusse and Varmus 2012 EMBO
Janda et al 2012 Science
Nusse and Varmus 2012 EMBO
Inestrosa and Arenas, 2009, Nat Rev Neurosci
Farias et al., 2009, J Biol Chem
Cuitino et al., 2010, J Neurosci
Technical Support: Juan Godoy and Inés Poblete
In collaboration with Dr. Francisco José Muñoz (Universidad Pompeau Fabra, Barcelona) and Dr. Juan Pablo García-Huidobro (P. Universidad Católica de Chile, CARE)
Condition Reversal potential
Control -53 ± 7 mV
Wnt-5a -56 ± 4 mV
TEA -55 ± 2 mV
-150 -100 -50 0 50 100 150
-500
0
500
1000
1500 Kv 3.2 control SNAP 100 nM SNAP 25 nM SNAP 1 M
mV
pA
-150 -100 -50 0 50 100 150
-500
0
500
1000
1500 Kv 3.2 control Wnt 1:100 Wnt 1.25 Wnt 1:10
mV
pA
-150 -100 -50 0 50 100 150
-500
0
500
1000
1500 Control Kv 3.2 SNAP Wnt-5a 7-NI
mV
pA
-150 -100 -50 0 50 100 150
-500
0
500
1000
1500
mV
pA
Control Kv 3.2 1 nM 10 nM 100 nM 1000 nM
HEK
Neuron
Control sFRP 7-NI Go 69760
200
400
600
800
*
*
* Wnt-5a
Mem
bra
ne
resi
sten
ce (
M)
Wnt-5a
¿Afecta la actividad miniatura?
Control AMPA NMDA0
20
40
60
80
100 Control 15 min
*
*
Am
plit
ud
e m
init
ure
cu
rren
t (p
A)
0
20
40
60
80
100
Formyl-6aaWnt-5a + sFRPWnt-5aControl
**
Am
plit
ud
e m
init
ure
cu
rren
t (p
A)
Control AMPA
Wnt-5a AMPA
Control NMDA
Wnt-5a NMDA
AMPA
NMDA
Control
Control
Wnt-5a
Wnt-5a
0,0
0,5
1,0
1,5
2,0
2,5
3,0
mN
MD
A fre
quen
cy (pA
)
**
Control Wnt-5a Wnt-5a + 7-NI
0
10
20
30
40
50
60
mN
MD
A a
mplit
ude
(pA
)
**
Wnt-5a + 7-NI
Wnt-5aControl
Plasticidad, efectos mas complejos del
ligando Wnt
-20 0 20 40 60 80 100 120 140 1600
1
2
3
4
5
fEPSP s
lope
% o
f C
ontr
ol (R
.U.)
Time (min)
-20 0 20 40 60 80 100 120 1400
2
fV (
%o
f C
on
tro
l)
X Axis Title
-40 -20 0 20 40 60 80 100 120
1,0
1,5
2,0
2,5
3,0
3,5
fEPSP s
lope
% o
f C
ontr
ol (
R.U
.)Time (min)
Control Wnt 5a ACSF
-60 -40 -20 0 20 40 60 80 100 1200
2
4
fEPSP s
lope
% o
f C
ontr
ol (R
.U.)
Time (min)
Wnt 5a 40 min Wnt 5a 20 min Control
-40 -20 0 20 40 600
1
2
3
4
5
Control 4 train TBS sFRP
fEPSP s
lope
(R.U
.)
Time (min)
-40 -20 0 20 40 600
1
2
3
4
5
fEPSP s
lope
(R.U
.)
Control Medium
Time (min)
-40 -20 0 20 40 600
1
2
3
4
5
Time (min)
fEPSP s
lope
(R.U
.)
Wnt-5a 2 train TBS Wnt-5a/sFRP sFRP
-20 0 20 40 600
1
2
3
4
5
Time (min)
fEPSP s
lope
(R.U
.) Control 7NI 0.5 M 7NI 1 M
-40 -20 0 20 40 600
1
2
3
4
5 Control 2 train 7NI
Time (min)
fEPSP s
lope
(R.U
.)
-20 0 20 40 600
1
2
3
4
5
Time (min)
fEPSP s
lope
(R.U
.)
Wnt-5a 2 train Wnt-5a 2 train/7-NI
0.0
0.5
1.0
1.5
2.0
2.5
3.0
Control Wnt-5a
20
mV
pu
lse
am
pli
tud
e (A
)
Control Wnt-5a
10 mM Calcium
BAPTA
BAPTA/Wnt-5a
0 mV
0 mV
20 mV
20 mV
BAPTA/Serum
BAPTA/Wnt-5a
Serum
0
200
400
600
800
1000
24 hrs
AT
P c
urre
nt (
nA)
Control Wnt-5a
Acute
0 1 2 3 4
0
20
40
60
80
100
120
Control Wnt-5a
Via
bili
da
d %
of
co
ntr
ol
Days
-200 -150 -100 -50 0 50 100
-1000
-800
-600
-400
-200
0
200
nA
mV
Control Wnt-5a 24 hrs Wnt-5a
-50
-40
-30
-20
-10
0
10
20
72 hrs48 hrs
Mem
bra
ne
pote
ntial
(m
V
Control Wnt-5a
24 hrs
500
nA
Control Prog. Wnt-5a
Control
Prog.
Wnt-5a
Medium
0 5 10 15 20 25
0
20
40
60
80
100
120
Control Medium Prog Wnt-5a
% G
VB
D
Hours of Washed
AgradecimientosDr. Nibaldo InestrosaDr. Fernando Romero, UFRODr. Ataulfo Martinez-Torres UNAMFelipe Serrano
Time (min)
0
0.5
1.0
1.5
2.0Control
ACSF
Case
Depleted
fEPS
P A
mplit
ude
(rela
tive u
nit
s)-10 0 10 20 30
Serum
Serum
Control
0
0.5
1.0
1.5
Facilit
ati
on
In
dex
R2
-R1
/R1
Case
0.5 m
V
50 ms
fEP
SP
A
mp
litu
de
(rela
tive
un
its)
ACSF
00.5
1.0
1.5
2.0
Control Case Depleted
*
0.5 m
V
10 ms
Serum
0
0.5
1.0
1.5
2.0CaseACSF
Depleted
Time (min)
fEPS
P A
mplit
ude
(rela
tive u
nit
s)-
100 10 20 30
CSF
CSF
ACSF 0
0.5
1.0
1.5
2.0
DepletedCase
CSF
fEPSP
Am
plit
ude
(rela
tive u
nit
s) *
0.5
m
V
10 ms
Control
0
0.5
1.0
1.5
CSF
Faci
litati
on Index
R2
-R1
/R1
0.5
m
V
50 ms
1.0
3.5
6.0
8.5 0.025 g/ml0.25 g/ml2.5 g/mlACSF
-10
Time (min)
100 403020 50
fEP
SP
Am
plitu
de(r
ela
tive
units
)
0
Anti-P
0
1.0
3.5
6.0
8.5
*
*
ACSF 2.5 g/ml
0.25 g/ml
0.025 g/ml
fEP
SP
Am
plitu
de(r
ela
tive
units
)
Anti-P
0.5 m
V
10 ms
-100
1.0
2.0
3.0
4.0Anti-P (0.25 g/ml)
ACSFAnti-P + APV (50 M)
Time (min)
fEPS
P A
mplit
ude
(rela
tive u
nit
s)
100 403020 50
ACSF
0
1.0
2.0
3.0
4.0
Anti-P Anti-P + d-APV
*
0.5
m
V
10 ms
fEPS
P A
mplit
ude
(rela
tive u
nit
s)
-20 -10 0 10 20 300
1
2
3
Time (min)
fEP
SP
am
plitu
de
(rela
tive u
nit
s)
KO mouse Control