TEMPLATE DESIGN © 2008
www.PosterPresentations.com
Motor Deficits in a Novel Optineurin Mouse for Amyotrophic Lateral Sclerosis
McKee, C.1, Bomze, H.1, Rhodes, T.2,3, Means, C.2,3, Rodriguiz, R.M.2,3, Wetsel, W.C.2,3,4, Tseng, H.C.1
1Department of Ophthalmology, 2Department of Psychiatry and Behavioral Sciences, 3Mouse Behavioral and Neuroendocrine Analysis Core Facility, 4Departments of Cell Biology and Neurobiology
Duke Eye Center and Duke University Medical Center, Durham, NC 27710
Introduction
Methods
Conclusions
Mutations in optineurin are associated with
familial glaucoma and amyotrophic lateral
sclerosis (ALS).
The pathophysiological mechanisms of both
neurodegenerative diseases are unknown.
Optineurin functions in autophagy, TNF-NFκB
signal transduction, protein trafficking, and
polyubiquitin binding.
We hypothesized that ALS-associated optineurin
mutations result in a loss-of-function disease
mechanism.
To test this, we generated an optineurin knockout
mouse and compared knockout animals to
C57BL/6J controls in motor behavioral tests.
This work is supported by a K12-EY016333, K08-
EY021520, and National Eye Institute core grant
P30EY005722 to the Duke Eye Center.
Acknowledgements
Using the Cre-LoxP system (Fig 1), “floxed”
optineurin mice were generated and backcrossed
to a C57BL/6J background for 5-6 generations.
When crossed to CMV-Cre, complete genetic
deletion of optineurin was confirmed by Western
blotting of dissected retinas (Fig. 1).
Motor/behavioral tests were performed using 3-4
month old optineurin homozygous knockout
(KO) and C57BL/6J (C57) mice.
Disclosures
The authors have no commercial interests in this work.
Results
Figure 1. Generation of optineurin
knockout (KO) mice
Lo
xP
Lo
xP
Cre
Lo
xP
Floxed OPTN
OPTN
Knockout
Exon 1
frt
frt NeoR
ATG
OPTN N-term
OPTN C-term
-/- +/- +/+
GAPDH
Although more physically active (open field
test), optineurin KO mice exhibited deficits in
balance and coordination (rotarod steady speed,
foot fault, and pole walk tests) compared to
control C57 mice.
Consistently, optineurin KO mice required
more time in stabilization gait phases (stance &
propel), spent less time in a destabilizing phase
(swing), and exhibited reduced paw print area
and angle.
Front paw grip was stronger in optineurin KO
compared to control C57 mice. The KO mice
also have weaker rear paws than front paws,
but were comparable to control mice.
Motor impairments observed in our optineurin
KO mice suggest they may be useful as animal
models to study ALS.
Diameter +/+ (C57) -/- (KO)
28 mm 0% 18.2%
18 mm 10% 63.6%a
12 mm 40% 36.4%
9 mm 30% 63.6%b
A p < 0.011, b p = 0.122
Figure 2. Open Field Tests
Figure 3. Rotarod Accelerating and Steady Speed Tests
Figure 4. Foot Fault Tests
Figure 5. Grip Strength Tests
Results Results Results
Figure 6. Pole Walk Tests
Figure 7. Forced Gait Analysis
Figure 8. Forced Gait Phases
Open Field Test Statistical Difference
Vertical Activity -/- (KO) showed more vertical postures
Repetitive Activity No difference
Lapping/ Circling No difference
Thigmotaxis No difference
Distance in center -/- (KO) more active In center
Time spent in center No difference
Percent of C57 (n=10) and KO (n=11) mice which
required more than 1 trial to walk across pole
Mean ± S.E.M, n = 10 for C57, n = 9 for KO * p < 0.05 by Repeated Measures Analyses of Variance (RMANOVA)
Mean ± S.E.M, n = 10 for C57, n = 9 for KO,
p = 0.077, ** p = 0.011
Repeated Measures Analyses of Variance (RMANOVA)
Mean ± S.E.M, n = 10 for C57, n = 9 for KO, * p = 0.005, ** p = 0.001,*** p = 0.023
Repeated Measures Analyses of Variance (RMANOVA)
* *
*
*
**
***
Mean ± S.E.M, n = 10 for C57, n = 9 for KO, * p < 0.001 by Repeated Measures Analyses of Variance (RMANOVA)
* *
** *
Mean ± S.E.M, n = 10 for C57, n = 11 for KO, * p = 0.003, ** p = 0.001
Repeated Measures Analyses of Variance (RMANOVA)
Mean ± S.E.M, n = 10 for C57, n = 9 for KO * p = 0 .03, ** p < 0.001
Repeated Measures Analyses of Variance (RMANOVA)
**
*
*
Mean ± S.E.M, n = 10 for C57, n = 9 for KO * p = 0.018, ** p = 0.009 by t-test
** *
*
**
Recommended
