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Gait Analysis in Laboratory Animals:
Studying Coordinated Movement and
Associated Disorders
Sponsored by:
Martin Hess
InsideScientific
Tom Hampton, PhD
Mouse Specifics Inc.
Carol Milligan, PhD
Wake Forest School of
Medicine
Charles Meshul, PhD
VA Medical Center/Portland
& Oregon Health & Science
University
InsideScientific is an online educational environment
designed for life science researchers. Our goal is to aid in
the sharing and distribution of scientific information
regarding innovative technologies, protocols, research
tools and laboratory services.
Thank you to our webinar sponsor
DigiGait is a widely published ventral plane videography instrumentation available for gait
analysis in laboratory animals. Voluntary and treadmill walking, DigiGait performs gait
analysis of mice and rats over a range of walking and running speeds. Learn More >>>
Gait analysis in animal models of
human diseases –
measures of strength, balance,
& coordination
Thomas Hampton, PhD
President and CSO
Mouse Specifics, Inc.
Copyright 2015 T. Hampton, Mouse Specifics, Inc. & InsideScientific. All Rights Reserved.
What is gait analysis?
Gait analysis is the
quantitative assessment of
the manner of movement, as
it applies to ambulation
[walking and running].
When is gait disturbed?
• Arthritis
• Bone fractures
• Traumatic Brain Injury
• Amyotrophic lateral sclerosis
• Ataxia
• Medulloblastoma
• Spinal cord injury
• Huntington’s disease
• Nerve injury
• Orthopedic injuries
• Parkinson’s disease
• Multiple sclerosis
• Stroke
• Muscular dystrophy
Amyotrophic lateral sclerosis
Dr. Carol Milligan
Parkinson’s disease
Dr. Charles Meshul
Why gait analysis?
Patients want to walk!
Why gait analysis?
Why treadmill gait analysis?
1. rodents rarely follow instructions
2. walking speed significantly impacts posture & kinematics
3. higher repeatability and lower standard error
4. explore challenge conditions to highlight subtle problems
5. early detection
What does gait analysis measure?
How long is my stride?
How far apart are my feet?
Do my toes point inwards?
Is my swing exaggerated?
How steady am I on my feet?
The DigiGait™ Imaging System, is patented
instrumentation for studying gait by
examining the ventral view of subjects as
they walk on a transparent treadmill belt.
A high speed camera
images the ventral view of
animals as they walk, run,
limp, or hop on the
motorized transparent
treadmill belt.
AI examines each paw to
determine its position
relative to the treadmill
belt, resulting in gait
signals for each limb.
Braking
Propulsion
Stance
Stride
Swing
Pawarea
Time
Multiple strides
Voluntary and
treadmill
Range of speeds
(0-100 cm/s)
Horizontal,
incline, & decline
How do my animals walk?
Stride length
Stance width
Cadence
Paw placement angle
Swing time
Step-to-step stability
Braking duration
Propulsion duration
Gait symmetry
Paw overlap
Paw area
Tau
Rate of loading
Sciatic Functional Index
……….
18 cm/s
36 cm/s
Guinea PigEven small differences
in walking speed
significantly impact
gait metrics, including
stride length and paw
angles.
Ensemble
averaged
gait signals:
note good
symmetry
between left &
right forelimbs
(top) & hind
limbs (bottom).
Sciatic nerve
injury
5
7
9
11
13
15
17
19
32
41
52
63
76
97
110
Life Day
Hindpawplacementangle(degrees)
Wild-type
SOD1 G93A
P<0.05
ALS model
PD model
5
7
9
11
13
15
17
19
32
41
52
63
76
97
110
Life Day
Hindpawplacementangle(degrees)
Wild-type
SOD1 G93A
Stride length changes in
mouse ~0.5 cm.
“ we can’t
just place a
banana in
front of a
mouse to
cause a
disturbance”
Changing the
properties of
the walking
surface
primarily
affected
forelimb
kinematics.
Oil applied to walking surface
Changing the
properties of
the walking
surface
primarily
affected
forelimb
kinematics.
Oil applied to walking surface
How do lab animals
manage obstacles in their
walking path?
How are obstacles managed?
How might neurotoxins affect ability to detect and
negotiate obstacles?
Baseline
Ethanol
Pnd 14 Pnd 25
Pnd 14
Study of developmental
abnormalities
• Cerebral palsy
• Muscular dystrophy
• Spinal muscular atrophy
 Huntington’s disease [3NP, R6/2]
 Parkinson’s disease
[MPTP, 6-hydroxydopamine]
 Amyotrophic lateral sclerosis
[TDP-43, SOD1 G93A]
 MS
 OA, RA, Pain
 Stroke
 Spinal Cord Injury
The DigiGait™ Imaging System is the most widely
published treadmill gait analysis system available.
Recent Publications:
 Sashindranath et al.....craniotomy and traumatic brain
injury. Behav Brain Res. 2015
 Poulet et al. ...articular cartilage lesions.
Osteoarthritis Cartilage. 2015
 Gadalla et al. Gait analysis in a Mecp2 knockout....
PLoS One. 2014
Benefits
In vivo
Easy-to-use
Turn-key solution
Scalable
Automated
Digital information
Relevant metrics of motion
Integrates strength, balance,
coordination
Replaces multiple motor
function tests
Visual record of study
 Traumatic Brain Injury
 ALS, HD, PD
 Pain
 Muscular dystrophy
 Multiple sclerosis
Neuropathy
 Drug screening
Features Benefits Applications
The DigiGait™ Imaging System has features that bring
benefits to numerous disease applications.
Behavioral Deficits Correlate
with Early Pathology in the
SOD1 mouse model of ALS
Carol Milligan, Ph.D.
Professor,
Department of
Neurobiology & Anatomy
Wake Forest School of Medicine
Copyright 2015 C. Milligan, Mouse Specifics Inc. & InsideScientific. All Rights Reserved.
a. Amyotrophic Lateral Sclerosis and the usefulness (or not)
of the SOD1G93A mouse as an experimental model system
b. The importance of thorough, scientific characterization of experimental
model systems
c. The importance of including behavioral assays in determining
pathological processes
What are we going to discuss?
Ron Oppenheim
James Caress
Tom Hampton
David Gifondorwa
Mac Robinson
Anna Taylor
David Prevette
Thomas Gould
Ramon Jimenez-Moreno
Sherry Vinsant
Carol Mansfield
Jane Strupe
Phonepasong Arounleut
Masaaki Yoshikawa
Vickie Moore
Acknowledgements
Funding:
Wake Forest Brian White Funds
Blazeman Foundation for ALS
NIH/NINDS R01NS069212
NIH/NINDS R01NS036081
Robert Packard Center for ALS
Research at Johns Hopkins
Collaborators and Colleagues:
350,000 individuals in the world have
ALS/MND
120,000 diagnosed each year - 100,000 will
die each year
30,000 individuals in US affected with 5000
new cases each year.
The ALS center at WFUBMC sees
approximately 150 patients/year.
Adult onset (+ 55 years)
5-10% of cases are familial (fALS) with
remaining 90-95% being sporadic (sALS).
Amyotrophic Lateral Sclerosis (ALS)
Motor Neuron Disease (MND)
http://keribstill.com/
Amyotrophic Lateral Sclerosis (ALS)
Motor Neuron Disease (MND)
Patients present with difficulties with:
• speech and swallowing
• muscle weakness and atrophy (loss of large
motoneurons of brainstem and spinal cord)
• muscle fasciculations/twitching
• hyperreflexia and/or spasticity
(lesions to upper motoneurons)
Intellect, cardiovascular and other body functions
are not affected.
With no substantially effective treatment, patients
die on average thee years after diagnosis.
http://keribstill.com/
90% sporadic
Variations in DCTN1 (dynactin 1), NEFH
(neurofilament, heavy polypeptide), PRPH
(peripherin) and SMN1 (survival motor neuron 1)
increase chances of developing ALS.
ALS Patients and Genetics
ALS Patients and Genetics
10% familial
C9orf72 – 30-40% sALS- normal gene has a GGGGCC stretch, in ALS
patients this can be repeated > 30 times
SOD1 – 20% - Superoxide Dismutase 1- mutation appears to be toxic
gain of function
TARDBP (TDP-43) - 5% - TAR DNA binding protein, binds to RNA and
ensures stability
FUS – 5% - fused in sarcoma – binds DNA and regulates transcription
ANG – 1% - angiogenin, ribonuclease involved in angiogenesis
ALS2, SeTX, VAPB
• Autosomal dominant missense point mutation in SOD1 identified
in a subset of familial ALS patients
(Rosen et al., 1993. Nature. 362(6415):59-62).
• 10-15% of familial ALS cases (1% of all cases) possess mutations in
Cu+2/Zn+2 superoxide dismutase (SOD).
• Cu+2/Zn+2 SOD1 catalyzes the conversion of .O2 to O2 and H2O2.
• Mutation is toxic “gain of function”
• Cytoplasmic superoxide dismutase
• Mutation suggest a role for free radical damage in ALS.
Genetics and ALS?
• Gurney et al., 1994.
Science 264(5166):1772-5
• Commercially available from Jackson
Laboratory
• Females do not breed
• Autosomal dominant
• Develop “symptoms” at about 90 days
• Age of symptom onset is determined
when mice can not fully extend legs.
• Die by about 120-140 days
G93AG93A mutant
SOD1 mouse
Wild-Type
The SOD1G93A mouse model of ALS has been used in
many pre-clinical studies.
a, Average age of symptom onset is significantly delayed in rhHsp70 injected animals (n = 7)
compared with Riluzole treated (n = 12) and untreated mice (n = 11; ***p ≤ 0.001).
Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80.
SOD1 mice exhibit denervation of the neuromuscular
junction (NMJ).
a, Injection of rhHsp70 resulted in an increased percentage of innervated NMJs in the MG at P90.
Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80.
Motoneurons die by endstage in the SOD1G93A mouse.
Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
Modified from
Tradi onal coun ng does not reveal loss of MNs un l ends
But the number of vacuolated MNs predicts MNs that will de
• Charcot J-M (1874). De la sclérose latérale amyotrophique.
Prog Med. 2: 325-327, 341-342, 453-455.
• Duchen LW, Strich SJ (1968). An hereditary motor neurone disease with progressive denervation
of muscle in the mouse: the mutant ‘wobbler.’ J. Neurol Neurosurg Psychiatry 31 (6): 535-42)
• Messer, A., and Flaherty, L. (1986). Autosomal dominance in a late onset motor neuron disease
in the mouse. J. Neurogenet 3: 345-355.
• Gurney ME, Pu H, Chiu AY, Dal Cnato MC et al., (1994). Motor neuron degeneration in mice that
express a human Cu,Zn superoxide dismutase mutation. Science 264(5166):1772-5.
• Wegorzewska I, Bell S, Cairns NJ, Miller TM, Baloh RH (2009). TDP-43 mutant transgenic mice
develop features of ALS and frontotemporal lobar degeneration. PNAS 106(44): 18809-14.
Landmark papers in ALS
• Charcot J-M (1874). De la sclérose latérale amyotrophique.
Prog Med. 2: 325-327, 341-342, 453-455.
• Duchen LW, Strich SJ (1968). An hereditary motor neurone disease with progressive denervation
of muscle in the mouse: the mutant ‘wobbler.’ J. Neurol Neurosurg Psychiatry 31 (6): 535-42)
• Messer, A., and Flaherty, L. (1986). Autosomal dominance in a late onset motor neuron disease
in the mouse. J. Neurogenet 3: 345-355.
• Gurney ME, Pu H, Chiu AY, Dal Cnato MC et al., (1994). Motor neuron degeneration in mice that
express a human Cu,Zn superoxide dismutase mutation. Science 264(5166):1772-5.
• Wegorzewska I, Bell S, Cairns NJ, Miller TM, Baloh RH (2009). TDP-43 mutant transgenic mice
develop features of ALS and frontotemporal lobar degeneration. PNAS 106(44): 18809-14.
Title…
When and where does ALS
pathology begin?
What cells and processes are
involved?
SOD1G93A High Expression Chronology of Pathophysiology
• MNs are hyperexcitable (Pieri 2003; Kuo 2004; De Vos 2007)
• Transient behavioral deficits (Van Zundert 2008; Saxena 2009)
• Golgi fragmentation, mitochondrial vacuolization and damage
(Bendotti 2001; Gould 2006; Martin 2007)
• ER stress (Saxena 2009)
• SOD1 aggregates (Johnston 2000; Turner 2003; Gould 2006)
• Reduced motor units and muscle denervation (Frey 2000; Pun 2006; Gould 2006;
Hegedus 2007)
• Axonal transport deficits (Ligon 2005)
• Motor deficits (Open field test, grip strength (Barneoud 1997; Ligon 2005; Wooley
2005;Haywoth 2009)
• Reduced numbers of spinal motoneurons, corticospinal and bulbar spinal neurons
(Zang 2002; Shin 2007; Martin 2007)
• Disrupted BBB (Zhong 2008)
• Glial activation (Gould 2006)
• VR and DR axon loss (Fischer 2004; Fischer 2005)
• Hindlimb tremor (Chiu 1995)
• Rotorod deficits (Fischer 2005)
• MN cell death (Chiu 1995; Gould 2006)
Progressive paralysis, loss of 50% MNs and death
P30
Birth
P60
P90
P120-endstage
Pre-symptomatic?
Pre-symptomatic
rhHsp70 treatment
delays symptom
onset in G93A mice
Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of
amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80.
• Treatment beginning at
postnatal day (P) 50 with
rhHsp70 significantly
delayed symptom onset
and extended survival
• Treatment beginning at
P90 had no effect on
symptom onset or
survival.
• Treatment beginning at
P30 had greatest effects
on extending survival.
wt
BaxKO
SOD1
SOD/
BaxKO1
P140
end‐
stage
Dissociation of
Motoneuron Death
from ALS
• Loss or pro-apoptotic protein
Bax prevents loss of MN
during normal development
• Loss of Bax in the SOD1
mouse prevents MN cell
death
• However, while there was a
slight extension of survival,
the BAX deficient SOD1 mice
still died.
• Denervation of NMJ still
occurs and in SOD1 mice,
denervation occurs quite
early.
When and where is earliest pathology?
Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
When and where is earliest pathology?
Tibialis Anterior (TA)
• Anterior (skin)- Type 2B muscle fibers
• Posterior (bone)- Type 2A, B , x muscle fibers
• Motoneurons that innervate type 2B fibers are
most susceptible in ALS/MND.
Soleus
• Type 1 muscle fibers
• Motoneurons that innervate type 1
fibers are less susceptible in ALS/MND.
Initial denervation of TA NMJs begins after postnatal day 14 and before day 30
Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
Abnormalities are
observed in the MN
soma in region of the
TA motor pool at P30
Wild Type
Wild Type
SOD+
SOD+
• Enlarged and vacuolated
mitochondria are
observed
• Clear, empty cytoplastmic
vacuoles are present
P30 distal dendrites
show more
profound changes.
• Large, vacuolated
mitochondria are
prominent
• Clear, empty cytoplasmic
vacuoles are also present
in distal dendrites
• Distal dendrites are where
many excitatory synapses
are found on MNs
Fewer but larger mitochondria are observed in motoneuron
soma at postnatal day 30.
Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
Mitochondria membrane potential, ATP content and
generation are reduced in SODG93A at day 30.
Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
SODG93A SODG93A
Mitochondrial abnormali es are seen in axons in TA and occasionally in
soleus at P30.
TA soleus
SOD1 SOD1
WT
Abnormal mitochondria are observed in axons at day 30.
Mitochondria morphological alterations are also observed in motoneuron
terminals in NMJs of TA and soleus muscles.
SOD1 WT
Mitochondria morphological alterations are also observed in motoneuron
terminals in NMJs of TA and soleus muscles.
SOD1 WT
Fewer but larger mitochondria are observed
presynaptic terminals in both TA and soleus NMJs.
• Using ultrastuctural characteristics, the
number and type of afferent synapses
was determined at day 30.
• A C-type synapse that is usually
excitatory is shown in A.
• A type 2 synapse that is usually
inhibitory is shown in B.
• A type 1 synapse that is usually
excitatory is shown in C.
• In the SOD1 animals, synaptic
morphologies appeared normal.
Are there other indicators of
pathology at day 30?
Wild Type
SOD1
• There is a decrease in the number of
excitatory, Type 1 synapses on the MN
cell body.
• There is an increase in the number of
C-type synapses on the MN cell body.
• This is also a decrease in the number of
excitatory, Type 1 synapses on distal
dendrites.
The number and type of
afferent synapses is altered
at day 30.
Axo-somatic synapses
Axo-dendritic synapses
• Mitochondrial abnormalities are abundant in dendrites,
soma and NMJ presynaptic terminal.
• Mitochondria membrane potential, ATP content and generation are reduced in
lumbar spinal cord.
• Small, empty cytoplasmic vacuoles appear in motoneuron soma and dendrites.
• Decrease in total synapse and excitatory synapses in distal dendrites
• No change in total number of synapses, but decrease in excitatory and increase in C-
terminals
• Denervation of tibialis anterior and medial gastrocnemius muscles
• No apparent change in retrograde transport.
By postnatal day 30 there are obvious pathological
changes observed in the SOD1 mouse.
Symptom onset in the SOD1 mouse occurs much later
than day 30.
Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80.
G93A
Wild-Type
P90
Age of symptom onset was
determined when mice can not
fully extend legs.
Observed typically day 90
The initial symptoms of ALS can be quite varied in different
people. One person may have trouble grasping a pen or lifting
a coffee cup, while another person may experience a change in
vocal pitch when speaking. ALS is typically a disease that
involves a gradual onset.
http://www.alsa.org/about-als/symptoms
Jon “the Blazeman” Blais (www.waronals.org)
He noticed he was having a hard time holding
things -- soap, toothbrush, etc. -- but he kept
putting off seeing someone about it.
We wanted to determine of the pathological changes were associated with symptom onset.
Gait dynamics were recorded using ventral plane videography using the Digigait Imaging
System.
Decreases in limb stance width and increases in variability of paw placement
angle occur at the time of initial denervation
Forelimb stance width
Hindlimb stance width
• In mouse models, pathological changes occur much
earlier than overt behavior changes.
• It is essential to characterize early pathology and utilize
appropriate behavioral analysis.
• By understanding the earliest pathology, we will
- gain insight into potential causes of disease,
- identify biomarkers, and
- develop effective therapeutics
Conclusions…
Changes in DigiGait measures in
a progressive animal model of
Parkinson's disease
Charles K. Meshul, Ph.D
Research Biologist, VA Medical
Center/Portland and Professor,
Department of Behavioral
Neuroscience
Oregon Health & Science University
Copyright 2015 C.K.Meshul, Mouse Specifics Inc. & InsideScientific. All Rights Reserved.
Acknowledgments
The important contributions from the following individuals are greatly appreciated:
Michelle Sconce
Madeline Churchill
Rebecca Hood
Natalie Goldberg
This research was supported by the Department of Veterans Affairs Merit Review Program
The Parkinson’s disease (PD) animal model
• Most PD rodent models primarily have used acute/subacute toxin administration, resulting
in >70-95% unilateral or bilateral loss of nigrostriatal dopamine. This results in significant
motor dysfunction.
• We have used a progressive loss of dopamine over a 4-week time period to try and simulate
the longer term loss of dopamine as seen in PD patients. This results in a 60-80% loss of
dopamine. This slower loss of dopamine most likely results in the brain adapting to the new
environment, making motor dysfunction more difficult to measure.
• Most treatment paradigms have used a neuroprotection strategy, which are not clinically
relevant. We have tested treatments either during (intervention) or following (restoration)
progressive MPTP administration.
Dopaminergic
and Behavioral
Correlations
• Increased dosing of MPTP
over a 4-week time period
results in a progressive
loss of dopamine.
• Treatment can be initiated
anytime during this 4-
week time period,
although the emphasis in
terms of translation is
more in restoration versus
protection.
Goldberg et al: Dopaminergic and Behavioral Correlates of Progressive Lesioning of the Nigrostriatal Pathway with 1-
Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine. Neuroscience 180:256-271 (2011).
Gait dynamics in vehicle and MPTP-treated (4mg/kg – 32 mg/kg)
at a walking speed of 24 cm/s
Goldberg et al.: Profiling changes in gait dynamics resulting from progressive MPTP-induced nigrostriatal lesioning. J. Neuroscience Research 89:1698-1706 (2011).
Sconce et al: Intervention with 7,8-dihydroxyflavone blocks further striatal terminal loss and restores motor deficits in a progressive mouse model of Parkinson’s disease. Neuroscience
290:454-471 (2015).
Intervention with 7,8-dihydroxyflavone (DHF): Gait changes
With MPTP treatment, the forepaws exerted a greater
distance from the midline compared to the
hindpaws (ie negative values).
Correlation of dopamine, sprouting and second messenger
biomarkers on measures of motor function
• With a decrease in TH/dopamine terminals within the striatum following MPTP treatment, there was an increase in
the time that the hind paws were on the belt during running. There was improvement following DHF treatment.
• Following DHF treatment, there was an increase in the levels of SCG 10 (sprouting factor) and the activated BDNF
receptor (pTrkB), resulting in gait symmetry between the forepaws and hindpaws.
Sconce et al: Intervention with 7,8-dihydroxyflavone blocks further striatal terminal loss and restores motor deficits in a progressive mouse model of Parkinson’s disease. Neuroscience
290:454-471 (2015).
.
Intervention
with Exercise
• Progressive treatment
with MPTP results in
changes in several
measures of gait, including
paw area, stance width
and shared stance,
suggesting instability due
to dopamine loss.
• Intervention with
voluntary exercise, starting
2 weeks after the start of
MPTP treatment, results in
recovery of these gait
measures.
Sconce et al: Intervention with exercise restores motor deficits but not nigrostriatal loss in a progressive MPTP mouse
model of Parkinson’s disease. Neuroscience 299:156-174 (2015).
Maximal rate of change of paw
area in contact with belt during
breaking phase.
% of stance of hind paws in contact
with belt at same time.
• With MPTP treatment,
levels of the glutamate
transporter increased,
which was positively
correlated with an
increase in paw area.
• Interestingly, as the
levels of the vesicular
dopamine transporter,
VMAT2, decreased, the
paw area increased.
Correlation of
dopamine and
glutamate biomarkers
Curcumin Treatment Improves Motor Behavior in ɑ-Synuclein Transgenic Mice
Spinelli et al: Curcumin treatment improves motor behavior in alpha-synulcein transgenic mice. Plos One, 10(6): e0128510 (2015).
MPTP: 4 weeks, followed by DHF treatment for 4 weeks (restoration). Most of the
DigiGait measures were significantly different 4 weeks after MPTP treatment (MPTP group)
compared to the 4wk MPTP only group.
Flavone-induced restoration of gait function following MPTP
Sconce et al: in preparation
• MPTP treatment resulted
in increased stance width,
decreased stance/swing,
increased swing time, and
increased paw angle.
• The changes in gait
described above were
reversed following DHF
treatment and these were
correlated with changes in
second messenger and
sprouting levels.
Correlation of sprouting
and second messenger
biomarkers with gait
measures following
flavone treatment.
Sconce et al: in preparation
Data courtesy of: Madeline Churchill (manuscript in preparation)
• 4 weeks of MPTP followed by 4 weeks of daily cyclosporin A treatment (restoration)
• Greater stance width implies overcompensation for gait instability; connected with step angle variance.
Effect of Cyclosporin A (CsA) treatment following MPTP
Blue bars = CsA treatment
White bars = vehicle treatment
Summary & Conclusions
1. Progressive loss of dopamine in a mouse model of PD, followed by withdrawal of
the toxin for 1-4 weeks, results in variable changes in several measures of gait. All
the MPTP-induced gait alterations appear to be connected to gait instability.
However, the slight differences in dopamine loss (60-75%) may contribute to
which gait measures are affected.
2. Although the same strain of mouse (male, C57Bl/6J, 12-15 weeks old) and dose
of MPTP are used for each study, the variability in the differences in gait
measures may simply replicate what is seen in patients with PD. Not all PD
patients develop exactly the identical motor disturbances. Our data may be
reflective of those differences seen in the human population despite the fact
that the same strain/sex/age of mouse is used.
Thank You!
For additional information on Mouse
Specifics equipment specially designed for
gait analysis in rodents please visit:
https://mousespecifics.com/

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Gait analysis in laboratory animals studying coordinated movement and associated disorders

  • 1. Gait Analysis in Laboratory Animals: Studying Coordinated Movement and Associated Disorders Sponsored by: Martin Hess InsideScientific Tom Hampton, PhD Mouse Specifics Inc. Carol Milligan, PhD Wake Forest School of Medicine Charles Meshul, PhD VA Medical Center/Portland & Oregon Health & Science University
  • 2. InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services.
  • 3. Thank you to our webinar sponsor DigiGait is a widely published ventral plane videography instrumentation available for gait analysis in laboratory animals. Voluntary and treadmill walking, DigiGait performs gait analysis of mice and rats over a range of walking and running speeds. Learn More >>>
  • 4. Gait analysis in animal models of human diseases – measures of strength, balance, & coordination Thomas Hampton, PhD President and CSO Mouse Specifics, Inc. Copyright 2015 T. Hampton, Mouse Specifics, Inc. & InsideScientific. All Rights Reserved.
  • 5. What is gait analysis? Gait analysis is the quantitative assessment of the manner of movement, as it applies to ambulation [walking and running].
  • 6. When is gait disturbed? • Arthritis • Bone fractures • Traumatic Brain Injury • Amyotrophic lateral sclerosis • Ataxia • Medulloblastoma • Spinal cord injury • Huntington’s disease • Nerve injury • Orthopedic injuries • Parkinson’s disease • Multiple sclerosis • Stroke • Muscular dystrophy Amyotrophic lateral sclerosis Dr. Carol Milligan Parkinson’s disease Dr. Charles Meshul
  • 8. Patients want to walk! Why gait analysis?
  • 9. Why treadmill gait analysis? 1. rodents rarely follow instructions 2. walking speed significantly impacts posture & kinematics 3. higher repeatability and lower standard error 4. explore challenge conditions to highlight subtle problems 5. early detection
  • 10. What does gait analysis measure? How long is my stride? How far apart are my feet? Do my toes point inwards? Is my swing exaggerated? How steady am I on my feet?
  • 11. The DigiGait™ Imaging System, is patented instrumentation for studying gait by examining the ventral view of subjects as they walk on a transparent treadmill belt.
  • 12. A high speed camera images the ventral view of animals as they walk, run, limp, or hop on the motorized transparent treadmill belt. AI examines each paw to determine its position relative to the treadmill belt, resulting in gait signals for each limb. Braking Propulsion Stance Stride Swing Pawarea Time
  • 13. Multiple strides Voluntary and treadmill Range of speeds (0-100 cm/s) Horizontal, incline, & decline
  • 14. How do my animals walk? Stride length Stance width Cadence Paw placement angle Swing time Step-to-step stability Braking duration Propulsion duration Gait symmetry Paw overlap Paw area Tau Rate of loading Sciatic Functional Index ……….
  • 15. 18 cm/s 36 cm/s Guinea PigEven small differences in walking speed significantly impact gait metrics, including stride length and paw angles.
  • 16.
  • 17. Ensemble averaged gait signals: note good symmetry between left & right forelimbs (top) & hind limbs (bottom).
  • 19. 5 7 9 11 13 15 17 19 32 41 52 63 76 97 110 Life Day Hindpawplacementangle(degrees) Wild-type SOD1 G93A P<0.05 ALS model PD model 5 7 9 11 13 15 17 19 32 41 52 63 76 97 110 Life Day Hindpawplacementangle(degrees) Wild-type SOD1 G93A Stride length changes in mouse ~0.5 cm.
  • 20. “ we can’t just place a banana in front of a mouse to cause a disturbance”
  • 21. Changing the properties of the walking surface primarily affected forelimb kinematics. Oil applied to walking surface
  • 22. Changing the properties of the walking surface primarily affected forelimb kinematics. Oil applied to walking surface
  • 23. How do lab animals manage obstacles in their walking path?
  • 24. How are obstacles managed?
  • 25. How might neurotoxins affect ability to detect and negotiate obstacles? Baseline Ethanol
  • 27. Pnd 14 Study of developmental abnormalities • Cerebral palsy • Muscular dystrophy • Spinal muscular atrophy
  • 28.  Huntington’s disease [3NP, R6/2]  Parkinson’s disease [MPTP, 6-hydroxydopamine]  Amyotrophic lateral sclerosis [TDP-43, SOD1 G93A]  MS  OA, RA, Pain  Stroke  Spinal Cord Injury The DigiGait™ Imaging System is the most widely published treadmill gait analysis system available. Recent Publications:  Sashindranath et al.....craniotomy and traumatic brain injury. Behav Brain Res. 2015  Poulet et al. ...articular cartilage lesions. Osteoarthritis Cartilage. 2015  Gadalla et al. Gait analysis in a Mecp2 knockout.... PLoS One. 2014
  • 29. Benefits In vivo Easy-to-use Turn-key solution Scalable Automated Digital information Relevant metrics of motion Integrates strength, balance, coordination Replaces multiple motor function tests Visual record of study  Traumatic Brain Injury  ALS, HD, PD  Pain  Muscular dystrophy  Multiple sclerosis Neuropathy  Drug screening Features Benefits Applications The DigiGait™ Imaging System has features that bring benefits to numerous disease applications.
  • 30. Behavioral Deficits Correlate with Early Pathology in the SOD1 mouse model of ALS Carol Milligan, Ph.D. Professor, Department of Neurobiology & Anatomy Wake Forest School of Medicine Copyright 2015 C. Milligan, Mouse Specifics Inc. & InsideScientific. All Rights Reserved.
  • 31. a. Amyotrophic Lateral Sclerosis and the usefulness (or not) of the SOD1G93A mouse as an experimental model system b. The importance of thorough, scientific characterization of experimental model systems c. The importance of including behavioral assays in determining pathological processes What are we going to discuss?
  • 32. Ron Oppenheim James Caress Tom Hampton David Gifondorwa Mac Robinson Anna Taylor David Prevette Thomas Gould Ramon Jimenez-Moreno Sherry Vinsant Carol Mansfield Jane Strupe Phonepasong Arounleut Masaaki Yoshikawa Vickie Moore Acknowledgements Funding: Wake Forest Brian White Funds Blazeman Foundation for ALS NIH/NINDS R01NS069212 NIH/NINDS R01NS036081 Robert Packard Center for ALS Research at Johns Hopkins Collaborators and Colleagues:
  • 33. 350,000 individuals in the world have ALS/MND 120,000 diagnosed each year - 100,000 will die each year 30,000 individuals in US affected with 5000 new cases each year. The ALS center at WFUBMC sees approximately 150 patients/year. Adult onset (+ 55 years) 5-10% of cases are familial (fALS) with remaining 90-95% being sporadic (sALS). Amyotrophic Lateral Sclerosis (ALS) Motor Neuron Disease (MND) http://keribstill.com/
  • 34. Amyotrophic Lateral Sclerosis (ALS) Motor Neuron Disease (MND) Patients present with difficulties with: • speech and swallowing • muscle weakness and atrophy (loss of large motoneurons of brainstem and spinal cord) • muscle fasciculations/twitching • hyperreflexia and/or spasticity (lesions to upper motoneurons) Intellect, cardiovascular and other body functions are not affected. With no substantially effective treatment, patients die on average thee years after diagnosis. http://keribstill.com/
  • 35. 90% sporadic Variations in DCTN1 (dynactin 1), NEFH (neurofilament, heavy polypeptide), PRPH (peripherin) and SMN1 (survival motor neuron 1) increase chances of developing ALS. ALS Patients and Genetics
  • 36. ALS Patients and Genetics 10% familial C9orf72 – 30-40% sALS- normal gene has a GGGGCC stretch, in ALS patients this can be repeated > 30 times SOD1 – 20% - Superoxide Dismutase 1- mutation appears to be toxic gain of function TARDBP (TDP-43) - 5% - TAR DNA binding protein, binds to RNA and ensures stability FUS – 5% - fused in sarcoma – binds DNA and regulates transcription ANG – 1% - angiogenin, ribonuclease involved in angiogenesis ALS2, SeTX, VAPB
  • 37. • Autosomal dominant missense point mutation in SOD1 identified in a subset of familial ALS patients (Rosen et al., 1993. Nature. 362(6415):59-62). • 10-15% of familial ALS cases (1% of all cases) possess mutations in Cu+2/Zn+2 superoxide dismutase (SOD). • Cu+2/Zn+2 SOD1 catalyzes the conversion of .O2 to O2 and H2O2. • Mutation is toxic “gain of function” • Cytoplasmic superoxide dismutase • Mutation suggest a role for free radical damage in ALS. Genetics and ALS?
  • 38. • Gurney et al., 1994. Science 264(5166):1772-5 • Commercially available from Jackson Laboratory • Females do not breed • Autosomal dominant • Develop “symptoms” at about 90 days • Age of symptom onset is determined when mice can not fully extend legs. • Die by about 120-140 days G93AG93A mutant SOD1 mouse Wild-Type
  • 39. The SOD1G93A mouse model of ALS has been used in many pre-clinical studies. a, Average age of symptom onset is significantly delayed in rhHsp70 injected animals (n = 7) compared with Riluzole treated (n = 12) and untreated mice (n = 11; ***p ≤ 0.001). Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80.
  • 40. SOD1 mice exhibit denervation of the neuromuscular junction (NMJ). a, Injection of rhHsp70 resulted in an increased percentage of innervated NMJs in the MG at P90. Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80.
  • 41. Motoneurons die by endstage in the SOD1G93A mouse. Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683 Modified from Tradi onal coun ng does not reveal loss of MNs un l ends But the number of vacuolated MNs predicts MNs that will de
  • 42. • Charcot J-M (1874). De la sclérose latérale amyotrophique. Prog Med. 2: 325-327, 341-342, 453-455. • Duchen LW, Strich SJ (1968). An hereditary motor neurone disease with progressive denervation of muscle in the mouse: the mutant ‘wobbler.’ J. Neurol Neurosurg Psychiatry 31 (6): 535-42) • Messer, A., and Flaherty, L. (1986). Autosomal dominance in a late onset motor neuron disease in the mouse. J. Neurogenet 3: 345-355. • Gurney ME, Pu H, Chiu AY, Dal Cnato MC et al., (1994). Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264(5166):1772-5. • Wegorzewska I, Bell S, Cairns NJ, Miller TM, Baloh RH (2009). TDP-43 mutant transgenic mice develop features of ALS and frontotemporal lobar degeneration. PNAS 106(44): 18809-14. Landmark papers in ALS
  • 43. • Charcot J-M (1874). De la sclérose latérale amyotrophique. Prog Med. 2: 325-327, 341-342, 453-455. • Duchen LW, Strich SJ (1968). An hereditary motor neurone disease with progressive denervation of muscle in the mouse: the mutant ‘wobbler.’ J. Neurol Neurosurg Psychiatry 31 (6): 535-42) • Messer, A., and Flaherty, L. (1986). Autosomal dominance in a late onset motor neuron disease in the mouse. J. Neurogenet 3: 345-355. • Gurney ME, Pu H, Chiu AY, Dal Cnato MC et al., (1994). Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264(5166):1772-5. • Wegorzewska I, Bell S, Cairns NJ, Miller TM, Baloh RH (2009). TDP-43 mutant transgenic mice develop features of ALS and frontotemporal lobar degeneration. PNAS 106(44): 18809-14. Title… When and where does ALS pathology begin? What cells and processes are involved?
  • 44. SOD1G93A High Expression Chronology of Pathophysiology • MNs are hyperexcitable (Pieri 2003; Kuo 2004; De Vos 2007) • Transient behavioral deficits (Van Zundert 2008; Saxena 2009) • Golgi fragmentation, mitochondrial vacuolization and damage (Bendotti 2001; Gould 2006; Martin 2007) • ER stress (Saxena 2009) • SOD1 aggregates (Johnston 2000; Turner 2003; Gould 2006) • Reduced motor units and muscle denervation (Frey 2000; Pun 2006; Gould 2006; Hegedus 2007) • Axonal transport deficits (Ligon 2005) • Motor deficits (Open field test, grip strength (Barneoud 1997; Ligon 2005; Wooley 2005;Haywoth 2009) • Reduced numbers of spinal motoneurons, corticospinal and bulbar spinal neurons (Zang 2002; Shin 2007; Martin 2007) • Disrupted BBB (Zhong 2008) • Glial activation (Gould 2006) • VR and DR axon loss (Fischer 2004; Fischer 2005) • Hindlimb tremor (Chiu 1995) • Rotorod deficits (Fischer 2005) • MN cell death (Chiu 1995; Gould 2006) Progressive paralysis, loss of 50% MNs and death P30 Birth P60 P90 P120-endstage Pre-symptomatic? Pre-symptomatic
  • 45. rhHsp70 treatment delays symptom onset in G93A mice Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80. • Treatment beginning at postnatal day (P) 50 with rhHsp70 significantly delayed symptom onset and extended survival • Treatment beginning at P90 had no effect on symptom onset or survival. • Treatment beginning at P30 had greatest effects on extending survival.
  • 46. wt BaxKO SOD1 SOD/ BaxKO1 P140 end‐ stage Dissociation of Motoneuron Death from ALS • Loss or pro-apoptotic protein Bax prevents loss of MN during normal development • Loss of Bax in the SOD1 mouse prevents MN cell death • However, while there was a slight extension of survival, the BAX deficient SOD1 mice still died. • Denervation of NMJ still occurs and in SOD1 mice, denervation occurs quite early.
  • 47. When and where is earliest pathology? Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
  • 48. When and where is earliest pathology? Tibialis Anterior (TA) • Anterior (skin)- Type 2B muscle fibers • Posterior (bone)- Type 2A, B , x muscle fibers • Motoneurons that innervate type 2B fibers are most susceptible in ALS/MND. Soleus • Type 1 muscle fibers • Motoneurons that innervate type 1 fibers are less susceptible in ALS/MND.
  • 49. Initial denervation of TA NMJs begins after postnatal day 14 and before day 30 Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
  • 50. Abnormalities are observed in the MN soma in region of the TA motor pool at P30 Wild Type Wild Type SOD+ SOD+ • Enlarged and vacuolated mitochondria are observed • Clear, empty cytoplastmic vacuoles are present
  • 51. P30 distal dendrites show more profound changes. • Large, vacuolated mitochondria are prominent • Clear, empty cytoplasmic vacuoles are also present in distal dendrites • Distal dendrites are where many excitatory synapses are found on MNs
  • 52. Fewer but larger mitochondria are observed in motoneuron soma at postnatal day 30. Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
  • 53. Mitochondria membrane potential, ATP content and generation are reduced in SODG93A at day 30. Vinsant S et al., (2013). Brain and Behavior 3 (4): 335–350. PMCID: PMC3869677 Vinsant S et al., (2013). Brain and Behavior 3 (4): 431–457. PMCID: PMC3869683
  • 54. SODG93A SODG93A Mitochondrial abnormali es are seen in axons in TA and occasionally in soleus at P30. TA soleus SOD1 SOD1 WT Abnormal mitochondria are observed in axons at day 30.
  • 55. Mitochondria morphological alterations are also observed in motoneuron terminals in NMJs of TA and soleus muscles. SOD1 WT
  • 56. Mitochondria morphological alterations are also observed in motoneuron terminals in NMJs of TA and soleus muscles. SOD1 WT Fewer but larger mitochondria are observed presynaptic terminals in both TA and soleus NMJs.
  • 57. • Using ultrastuctural characteristics, the number and type of afferent synapses was determined at day 30. • A C-type synapse that is usually excitatory is shown in A. • A type 2 synapse that is usually inhibitory is shown in B. • A type 1 synapse that is usually excitatory is shown in C. • In the SOD1 animals, synaptic morphologies appeared normal. Are there other indicators of pathology at day 30? Wild Type SOD1
  • 58. • There is a decrease in the number of excitatory, Type 1 synapses on the MN cell body. • There is an increase in the number of C-type synapses on the MN cell body. • This is also a decrease in the number of excitatory, Type 1 synapses on distal dendrites. The number and type of afferent synapses is altered at day 30. Axo-somatic synapses Axo-dendritic synapses
  • 59. • Mitochondrial abnormalities are abundant in dendrites, soma and NMJ presynaptic terminal. • Mitochondria membrane potential, ATP content and generation are reduced in lumbar spinal cord. • Small, empty cytoplasmic vacuoles appear in motoneuron soma and dendrites. • Decrease in total synapse and excitatory synapses in distal dendrites • No change in total number of synapses, but decrease in excitatory and increase in C- terminals • Denervation of tibialis anterior and medial gastrocnemius muscles • No apparent change in retrograde transport. By postnatal day 30 there are obvious pathological changes observed in the SOD1 mouse.
  • 60. Symptom onset in the SOD1 mouse occurs much later than day 30. Gifondorwa DJ, et. al. Exogenous delivery of heat shock protein 70 increases lifespan in a mouse model of amyotrophic lateral sclerosis. J Neurosci. 2007 Nov 28;27(48):13173-80. G93A Wild-Type P90
  • 61. Age of symptom onset was determined when mice can not fully extend legs. Observed typically day 90 The initial symptoms of ALS can be quite varied in different people. One person may have trouble grasping a pen or lifting a coffee cup, while another person may experience a change in vocal pitch when speaking. ALS is typically a disease that involves a gradual onset. http://www.alsa.org/about-als/symptoms Jon “the Blazeman” Blais (www.waronals.org) He noticed he was having a hard time holding things -- soap, toothbrush, etc. -- but he kept putting off seeing someone about it.
  • 62. We wanted to determine of the pathological changes were associated with symptom onset.
  • 63. Gait dynamics were recorded using ventral plane videography using the Digigait Imaging System.
  • 64. Decreases in limb stance width and increases in variability of paw placement angle occur at the time of initial denervation Forelimb stance width Hindlimb stance width
  • 65. • In mouse models, pathological changes occur much earlier than overt behavior changes. • It is essential to characterize early pathology and utilize appropriate behavioral analysis. • By understanding the earliest pathology, we will - gain insight into potential causes of disease, - identify biomarkers, and - develop effective therapeutics Conclusions…
  • 66. Changes in DigiGait measures in a progressive animal model of Parkinson's disease Charles K. Meshul, Ph.D Research Biologist, VA Medical Center/Portland and Professor, Department of Behavioral Neuroscience Oregon Health & Science University Copyright 2015 C.K.Meshul, Mouse Specifics Inc. & InsideScientific. All Rights Reserved.
  • 67. Acknowledgments The important contributions from the following individuals are greatly appreciated: Michelle Sconce Madeline Churchill Rebecca Hood Natalie Goldberg This research was supported by the Department of Veterans Affairs Merit Review Program
  • 68. The Parkinson’s disease (PD) animal model • Most PD rodent models primarily have used acute/subacute toxin administration, resulting in >70-95% unilateral or bilateral loss of nigrostriatal dopamine. This results in significant motor dysfunction. • We have used a progressive loss of dopamine over a 4-week time period to try and simulate the longer term loss of dopamine as seen in PD patients. This results in a 60-80% loss of dopamine. This slower loss of dopamine most likely results in the brain adapting to the new environment, making motor dysfunction more difficult to measure. • Most treatment paradigms have used a neuroprotection strategy, which are not clinically relevant. We have tested treatments either during (intervention) or following (restoration) progressive MPTP administration.
  • 69. Dopaminergic and Behavioral Correlations • Increased dosing of MPTP over a 4-week time period results in a progressive loss of dopamine. • Treatment can be initiated anytime during this 4- week time period, although the emphasis in terms of translation is more in restoration versus protection. Goldberg et al: Dopaminergic and Behavioral Correlates of Progressive Lesioning of the Nigrostriatal Pathway with 1- Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine. Neuroscience 180:256-271 (2011).
  • 70. Gait dynamics in vehicle and MPTP-treated (4mg/kg – 32 mg/kg) at a walking speed of 24 cm/s Goldberg et al.: Profiling changes in gait dynamics resulting from progressive MPTP-induced nigrostriatal lesioning. J. Neuroscience Research 89:1698-1706 (2011).
  • 71. Sconce et al: Intervention with 7,8-dihydroxyflavone blocks further striatal terminal loss and restores motor deficits in a progressive mouse model of Parkinson’s disease. Neuroscience 290:454-471 (2015). Intervention with 7,8-dihydroxyflavone (DHF): Gait changes With MPTP treatment, the forepaws exerted a greater distance from the midline compared to the hindpaws (ie negative values).
  • 72. Correlation of dopamine, sprouting and second messenger biomarkers on measures of motor function • With a decrease in TH/dopamine terminals within the striatum following MPTP treatment, there was an increase in the time that the hind paws were on the belt during running. There was improvement following DHF treatment. • Following DHF treatment, there was an increase in the levels of SCG 10 (sprouting factor) and the activated BDNF receptor (pTrkB), resulting in gait symmetry between the forepaws and hindpaws. Sconce et al: Intervention with 7,8-dihydroxyflavone blocks further striatal terminal loss and restores motor deficits in a progressive mouse model of Parkinson’s disease. Neuroscience 290:454-471 (2015). .
  • 73. Intervention with Exercise • Progressive treatment with MPTP results in changes in several measures of gait, including paw area, stance width and shared stance, suggesting instability due to dopamine loss. • Intervention with voluntary exercise, starting 2 weeks after the start of MPTP treatment, results in recovery of these gait measures. Sconce et al: Intervention with exercise restores motor deficits but not nigrostriatal loss in a progressive MPTP mouse model of Parkinson’s disease. Neuroscience 299:156-174 (2015). Maximal rate of change of paw area in contact with belt during breaking phase. % of stance of hind paws in contact with belt at same time.
  • 74. • With MPTP treatment, levels of the glutamate transporter increased, which was positively correlated with an increase in paw area. • Interestingly, as the levels of the vesicular dopamine transporter, VMAT2, decreased, the paw area increased. Correlation of dopamine and glutamate biomarkers
  • 75. Curcumin Treatment Improves Motor Behavior in ɑ-Synuclein Transgenic Mice Spinelli et al: Curcumin treatment improves motor behavior in alpha-synulcein transgenic mice. Plos One, 10(6): e0128510 (2015).
  • 76. MPTP: 4 weeks, followed by DHF treatment for 4 weeks (restoration). Most of the DigiGait measures were significantly different 4 weeks after MPTP treatment (MPTP group) compared to the 4wk MPTP only group. Flavone-induced restoration of gait function following MPTP Sconce et al: in preparation
  • 77. • MPTP treatment resulted in increased stance width, decreased stance/swing, increased swing time, and increased paw angle. • The changes in gait described above were reversed following DHF treatment and these were correlated with changes in second messenger and sprouting levels. Correlation of sprouting and second messenger biomarkers with gait measures following flavone treatment. Sconce et al: in preparation
  • 78. Data courtesy of: Madeline Churchill (manuscript in preparation) • 4 weeks of MPTP followed by 4 weeks of daily cyclosporin A treatment (restoration) • Greater stance width implies overcompensation for gait instability; connected with step angle variance. Effect of Cyclosporin A (CsA) treatment following MPTP Blue bars = CsA treatment White bars = vehicle treatment
  • 79. Summary & Conclusions 1. Progressive loss of dopamine in a mouse model of PD, followed by withdrawal of the toxin for 1-4 weeks, results in variable changes in several measures of gait. All the MPTP-induced gait alterations appear to be connected to gait instability. However, the slight differences in dopamine loss (60-75%) may contribute to which gait measures are affected. 2. Although the same strain of mouse (male, C57Bl/6J, 12-15 weeks old) and dose of MPTP are used for each study, the variability in the differences in gait measures may simply replicate what is seen in patients with PD. Not all PD patients develop exactly the identical motor disturbances. Our data may be reflective of those differences seen in the human population despite the fact that the same strain/sex/age of mouse is used.
  • 80. Thank You! For additional information on Mouse Specifics equipment specially designed for gait analysis in rodents please visit: https://mousespecifics.com/