The document summarizes research on the effects of Wnt ligands on synaptic plasticity. It discusses how Wnt-5a modulates potassium channel activity and miniature synaptic currents. Wnt-5a increases AMPA miniature currents but does not affect NMDA miniature currents. Wnt-5a enhances long-term potentiation induced by theta burst stimulation in the hippocampus in a manner dependent on calcium influx and nitric oxide signaling. Wnt-5a also increases neuronal viability and mitochondrial ATP production. The findings suggest Wnt ligands play complex roles in synaptic plasticity.

1. “Ligandos Wnt como neurotrofinas, plasticidad
sináptica modulada por ellos”
Dr. Jorge Parodi
Universidad Católica Temuco

3. Bubnik 2007 TRENDS

4. Schulte 2010, Phar- Review

5. Janda et al 2012 Science

6. Nusse and Varmus 2012 EMBO

7. Janda et al 2012 Science

10. Nusse and Varmus 2012 EMBO

11. Inestrosa and Arenas, 2009, Nat Rev Neurosci

12. Farias et al., 2009, J Biol Chem
Cuitino et al., 2010, J Neurosci

13. In collaboration with Dr. Francisco José Muñoz (Universidad Pompeau Fabra, Barcelona)
and Dr. Juan Pablo García-Huidobro (P. Universidad Católica de Chile, CARE)
Technical Support: Juan Godoy and Inés Poblete

15. Condition Reversal
potential
Control -53 ± 7 mV
Wnt-5a -56 ± 4 mV
TEA -55 ± 2 mV

17. 1500
Control Kv 3.2 1500
pA
HEK
Control Kv 3.2 pA 1 nM
SNAP
Wnt-5a
10 nM
7-NI 100 nM
1000 nM 1000
1000
500 500
mV mV
0 0
-150 -100 -50 0 50 100 150 -150 -100 -50 0 50 100 150
-500 -500
1500 1500
pA pA
Kv 3.2 control Kv 3.2 control
Wnt 1:100 SNAP 100 nM
Wnt 1.25 SNAP 25 nM
Wnt 1:10 SNAP 1 µM
1000 1000
500 500
mV mV
0 0
-150 -100 -50 0 50 100 150 -150 -100 -50 0 50 100 150
-500 -500
Neuron

18. 800
* Wnt-5a
Membrane resistence (MΩ )
600 *
400
* ¿Afecta la actividad
200
miniatura?
0
Control Wnt-5a sFRP 7-NI Go 6976

20. Control AMPA AMPA
100
* *
Amplitude miniture current (pA)
Control Wnt-5a
Wnt-5a AMPA 80
60
40
Control NMDA NMDA
20
Wnt-5a NMDA Control 0
Control Wnt-5a Wnt-5a + sFRP Formyl-6aa
Wnt-5a
100
Control *
Amplitude miniture current (pA)
15 min
80
*
60
40
20
0
Control AMPA NMDA

21. 3,0
m NMD A frequ en cy (p A )
2,5
**
2,0
1,5
1,0
0,5
0,0
Control Wnt-5a Wnt-5a
+ 7-NI
60
**
m NMD A am p litud e (pA )
50
40
30
20
10
0
Control Wnt-5a Wnt-5a
+ 7-NI

23. Plasticidad, efectos
mas complejos del
ligando Wnt

27. 2
5
fV (%of Control)
fEPSP s lop e % o f C on tro l (R .U.)
4
0
-20 0 20 40 60 80 100 120 140
X Axis Title
3
2
1
0
-20 0 20 40 60 80 100 120 140 160
Time (min)

28. 4 3,5
Control
fEPSP s lop e % of C on trol (R .U.)
fEPSP s lo pe % o f C o n trol (R .U.)
Wnt 5a
ACSF
3,0
2,5
2
2,0
Wnt 5a 40 min 1,5
Wnt 5a 20 min
Control
1,0
0
-60 -40 -20 0 20 40 60 80 100 120 -40 -20 0 20 40 60 80 100 120
Time (min) Time (min)

29. 5
5
Control
Medium
4
Control 4 train TBS
fEPSP s lo p e (R .U.)
sFRP
3
4 2
fEPSP s lop e (R .U.)
1
0
-40 -20 0 20 40 60
Time (min)
3
2
1
0
-40 -20 0 20 40 60
Time (min)

30. 5
Wnt-5a 2 train TBS
Wnt-5a/sFRP
4 sFRP
fEPSP s lope (R .U.)
3
2
1
0
-40 -20 0 20 40 60
Time (min)

31. 5 5
Control 2 train
Control 7NI
7NI 0.5 µ M
4 4
7NI 1 µ M
fEPSP s lop e (R .U.)
fEPSP s lo pe (R .U.)
3 3
2 2
1 1
0 0
-20 0 20 40 60 -40 -20 0 20 40 60
Time (min) Time (min)
5
Wnt-5a 2 train
Wnt-5a 2 train/7-NI
4
fEPS P s lope (R .U.)
3
2
1
0
-20 0 20 40 60
Time (min)

40. 3.0
20 mV pulse amplitude (µ A)
2.5
10 mM Calcium
2.0
1.5
1.0
0.5
0.0
Control Wnt-5a Control Wnt-5a

41. Serum BAPTA
20 mV
0 mV
BAPTA/Serum BAPTA/Wnt-5a
20 mV
0 mV
BAPTA/Wnt-5a

42. 120
Viabilidad % of control
100
80
60
40
Control
20 Wnt-5a
0
0 1 2 3 4
Days
1000
Control
Wnt-5a
800
ATP current (nA)
600
400
200
0
Acute 24 hrs

43. 20 Control
Wnt-5a
Mem b rane potential (m V )
10
0
24 hrs 48 hrs 72 hrs
-10
-20
-30
-40
-50 200
mV
0
-200 -150 -100 -50 0 50 100
-200
Control
-400 Wnt-5a
500 nA
24 hrs Wnt-5a
-600
-800
-1000
nA

44. Control Prog. Wnt-5a
Wnt-5a 120
Medium 100
80
% GVBD
60
40
Control
Medium
Prog. 20
Prog
Wnt-5a
0
Control 0 5 10 15 20 25
Hours of Washed

45. Agradecimientos
Dr. Nibaldo Inestrosa
Dr. Fernando Romero, UFRO
Dr. Ataulfo Martinez-Torres UNAM
Felipe Serrano

48. 2.0 Case
Control Serum
Depleted
1.5 ACSF
fEPSP Amplitude
(relative units)
1.0
0.5
Serum
0
-10 0 10 20 30
Time (min)
2.0
1.5
0.5 mV
0.5 mV
* 10 ms 50 ms
(relative units)
Facilitation Index
Amplitude
1.5
1.0
R2-R1/R1
fEPSP
1.0 0.5
0.5
0
ACSF Control Case Depleted
0
Serum Control Case

49. 2.0 ACSF
Case
CSF
fEPSP Amplitude
Depleted
(relative units)
1.5
1.0
0.5
CSF
0
- 0 10 20 30
10 Time (min)
2.0
mV
0.5
1.5
fEPSP Amplitude
mV
0.5
* 10 ms
(relative units)
Facilitation Index
1.5 50 ms
1.0
R2-R1/R1
1.0 0.5
0.5
0
ACSF Case Depleted
0
Control CSF
CSF

50. 8.5 0.025 µg/ml
0.25 µg/ml Anti-P
2.5 µg/ml
ACSF
fEPSP Amplitude
6.0
(relative units)
3.5
1.0
0
-10 0 10 20 30 40 50
Time
8.5 (min)
0.5 mV
* 10 ms
fEPSP Amplitude
(relative units)
6.0
3.5
*
1.0
0
ACSF 0.025 0.25 2.5
µg/ml µg/ml µg/ml
Anti-P

51. 4.0
Anti-P (0.25 µg/ml)
Anti-P + APV (50 µM)
ACSF
3.0
fEPSP Amplitude
(relative units)
2.0
1.0
0
-10 0 10 20 30 40 50
Time (min)
mV
0.5
4.0
*
10 ms
fEPSP Amplitude
(relative units)
3.0
2.0
1.0
0 ACSF Anti-P Anti-P +
d-APV

52. 3
KO mouse
Control
fEPSP amplitude
(relative units)
2
1
0
-20 -10 0 10 20 30
Time (min)