1. Focus on the Locus:
LC damage and NAergic treatment in TgAPP mice
Douglas L Feinstein
Dept of Anesthesiology
University of Illinois, Chicago
Jesse Brown VA Medical Center
December 16th, 2010

2. In the CNS, reductions of NA can cause:
Increased inflammatory responses
Increased amyloid burden
Reduced levels of neurotrophic factors (NGF, BDNF)
Reduced BBB integrity
Reduced neural progenitor cell maturation

3. In rodents, LC lession:
Reduces CNS NA
Increases inflammation, neuronal damage
Disrupts BBB
Reduces phagocytosis of amyloid
Increases amyloid burden, cognitive deficits

4. Evidence for LC damage in TgAPP mice
1. Atrophy of TH+ neurons occurs in aged V717F mice
German et al. 2005
2. Loss of TH+ neurons in aged PS:APP mice
O’Neil et al. 2007
3. Hippocampal and cortical NA levels are reduced in PS:APP mice
Pugh et al. 2007
4. NA transporter (NET-1) levels are reduced in PS:APP mice
Jardanhazi-Kurutz et al. 2010

5. Hypothesis:
Increasing CNS NA levels will provide benefit in
TgAPP mice with endogenous LC damage
5xFAD mice (R. Vassar): 3 mutations in hAPP; 2 mutations in PS1
Rapid Aβ accumulation and plaque appearance (6-8 weeks)
Neuronal damage; glial inflammation
Intraneuronal aggregated Aβ1-42
Cognitive deficits by 5-6 months

6. LC damage / inflammation in 5xFAD mice
A WT
LC
GFAP+ cells / field
140 *
120
100
TH GFAP 80
60
B 5xFAD 40
20
0
WT 5xFAD
TH GFAP

7. LC inflammation is not associated with plaques
A B
ThioS GFAP GFAP

8. LC TH+ cells are stressed in 5xFAD
Cell numbers Cell size
140 600
TH cell size (µm2)
*
TH+ cells / field
120 500
100
400
80
300
60
200
40
20 100
0 0
WT 5xFAD WT 5xFAD

9. L-DOPS (Droxidopa), a precursor of NA
L-DOPS
Tyrosine Hydroxylase OH
Aromatic amino acid L-AAAD
Decarboxylase
L-AAAD
Dopamine β-hydroxylase

10. L-DOPS / Droxidopa
Currently in phase III trials for the treatment
of Neurogenic Orthostatic Hypotension
Also provides benefit in congenital deficiency of
DBH
BBB permeable
Manufactured by Chelsea Therapeutics in the US;
Sumitomo in Japan

11. Treatment protocol
4.5 month old male 5xFAD mice (robust Aβ deposition)
Treated 3x / week for 4 weeks:
L-DOPS 200 mg/kg s.c.
Carbidopa 125 mg/kg i.p.
Atomoxetine (NA reuptake inhibitor) 20 mg/kg i.p.

12. Effects in Morris Water Maze
Vehicle DOPS
Latency to quadreant
60 *
50
40
30
20
10
0
Challenge Challenge
Re-challenge Re-challenge
After 1 week
1 day after 12 trial training in MWM, both vehicle and DOPS treated
mice responded comparably in a ‘challenge’ test. However 1 week later
the vehicle mice, but not the DOPS treated mice, showed a significant
reduced ability to find correct quadrant

13. L-DOPS increases CNS NA levels
[NA], pg/mg wet wt
500
*
400
300
200
100
0
Control DOPS

14. L-DOPS reduces Thioflavin S plaques and is
inversely correlated to NA levels in HC
Hippocampus Frontal Cortex
200 100 Control
TS plaques / field
Control
TS plaques / field
L-DOPS
150
L-DOPS 80
60
100
40
50
20
P = 0.0061 P = 0.1504
0 0
0 200 400 600 800 0 200 400 600 800
[NA], pg/mg [NA], pg/mg

15. L-DOPS neurotrophin mRNA expression
2.00
Ratio mRNA (DOPS:Ctrl)
* *
1.50
* * * *
1.00
*
*
0.50
0.00

16. LC:NA signaling is also perturbed in MS and
its rodent model EAE
Simonini et al. (2010) ASN Neuro
Polak et al (2011) Brain, in press

17. LC damage and inflammation is present in EAE
Control EAE
d d
c
c
v
v
SCD
SCD
TH GFAP TH GFAP

18. LC damage and inflammation occurs in MS
Increased GFAP staining
LC DTg
A Control TH GFAP
Area covered (% field)
5.00
* *
4.00
DTg 3.00
2.00
1.00
0.00
Control MS Control MS
B MS TH GFAP
TH Neuronal hypertrophy
0.35
% cells in size bin
∗
0.30
∗
0.25
0.20
DTg MS
0.15
0.10
0.05
HC
0.00
0 500 900 1300 1700 2100 2500
Size bin (µm2)

19. L-DOPS plus a NARI reduces clinical scores in EAE
Clinical Score (mean +- se)
3.0
2.5 Control, n=10
2.0
1.5
*
1.0 **
DOPS/Atom, n=9
0.5
0.0
0 10 20 30 40
Days post MOG booster
P= .0003 1 way Anova
*, p < .05 **, P<0.01 vs day 17

20. Conclusions
LC neuronal stress and inflammation occur in the 5xFAD
mouse model of AD
Increasing CNS NA levels provides cognitive and
pathological benefit in 5xFAD mice
Effects on plaque burden may involve increased NEP/IDE
Cognitive effects may involve increases in BDNF/Arc1
Dysfunction in the LC:NA system may be a feature of
several neurological disorders

21. Acknowledgements
Sergey Kalinin
Paul Polak
Alzheimer’s Association
National MS Society
NINDS, NIH
VA Merit Grant