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Automated Quantitative Measures of Forelimb Function in Rats and Mice
A webinar for researchers interested in transitioning from traditional qualitative forelimb assessments for rodents to quantitative measurements with automated, high-throughput systems.
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.
Automated Quantitative Measures of Forelimb Function in Rats and Mice
Copyright 2016 D. Sloan, Vulintus and InsideScientific. All Rights Reserved.
Drew Sloan, PhDChief Operating Officer,Vulintus, Inc.
What we will cover today…
• Automated Forelimb Assessments
• MotoTrak System Overview
• Walkthough of Typical Automated Assessment
1. Isometric Pull Task
2. Pronation/Supination Task
3. Lever Press Task
• Automated Forelimb Assessments for Preclinical Translation (Dr. Hays)
Common Forelimb Assessments
Pellet Retrieval Staircase Reaching
Pasta Matrix/Handling
Cylinder Ladder/Rope Walking
Grip Strength
Images credits: Tennant & Jones, 2009; Schaar et al., 2010; Smith et al., 2008
Common Forelimb Assessments
Automated Forelimb Assessments
Limitations
• Supervision-intensive
• Low subject throughput
• Manual scoring (requiring experimenter training)
• Relatively few trials per test
• Delay between behavior and score
Advantages
• Quantitative, objective measures
• High trial counts
• Immediate/real-time analysis
• Little experimenter training
• High subject-to-experimenter ratio
• Replicability across similar conditions
Limitations
• High equipment cost
• Simplified parameterization
Advantages
• Low equipment cost
• Complex parameterization
Automated Forelimb Assessments
Supination Methods Paper
Isometric Pull Methods Paper
Lever Press Methods Paper
The History of MotoTrak…
• Originally developed at UT-Dallas
• Vulintus created to license and commercialize
• NINDS Fast-Track Small Business grant 2014-2016
• Originally developed for rats, now also used with mice
• Used in 16 research papers and counting.• Published models: stroke, traumatic brain injury, spinal cord
injury, motor learning
• Coming soon: peripheral nerve injury
MotoTrakKey Features
Interchangeability
• Task modules
• Right-, left-, and either-paw configurations
Consistency Fail-Safes
• Automated manipulandumpositioning
• Enforced file organization
Real-Time Analysis
Validation
Pull Rat Stroke
Pull Rat SCI
Pull Mouse Stroke Pull Rat TBI
Lever Rat Stroke Knob Rat Stroke
Sloan et al., 2015
Becker et al., 2015
Pruitt et al., 2014
Ganzer et al., 2016
Hays et al., 2014
Meyers et al., 2016
Cross-Validation
• Isometric force vs skilled pellet retrieval
Sloan et al., 2015
• Isometric force vs pasta matrix
Sloan et al., 2015
• Lever press vs ladder rung
Unpublished
Isometric Pull Task – Objective
1 subject, 1 session data from Sloan et al., 2015
Isometric Pull Task – Mouse Models
• Modified Setup• Adjusted task dimensions• Smaller manipulandum• More sensitive force sensor
• Liquid reward vs. pellet reward
Isometric Pull Task - Shaping/Training Stages
• Training is organized into stages
• Variable stage parameters include:
1. hit criteria (pull force most common)
2. module position
3. reward schedule
4. auditory feedback
• Parameters can adaptively vary within a stage
Isometric Pull Task –Initial Shaping
• Any interaction with the handle is rewarded
• Pellets or other food can be used to initiate first interactions
Courtesy of Rennaker Lab, UT-Dallas
Isometric Pull Task –Typical Training
• Two 30-minute sessions each day
• 2+ hours between sessions
• 5 days per week
• Free feed on weekends
• Adaptive force threshold during session
• Maintains 50% reward schedule
• Maximum threshold ceiling
• Average ~15 days
N = 13, data from Sloan et al., 2015
Isometric Pull Task –Example Criterion
• Hit criterion: ≥ 120 gm of pull force within 2 seconds of initiation
• 20 gm for mice
• 5 consecutive days of >85% performance
• 250-300 trials per day
Isometric Pull Task –Post-Impairment
• Home cage recovery for 1-5 weeks
• Adaptive thresholding to maintain engagement with severe impairment
Isometric Pull Task –Analysis
• MATLAB-based (not required)
• Open-source
• Graphical Interface
• From individual trials to group analysis
Isometric Pull Task –Analysis
• MATLAB-based (not required)
• Open-source
• Graphical Interface
• From individual trials to group analysis
Supination Task – Objective
1 subject, 1 session data from Meyers et al., 2016
Supination Task – Training & Testing
• Shaping• Reward any interaction with free-spinning knob
• Training• Adaptive turn angle threshold• Increasing counterweight• More training stages (i.e. smaller steps) between shaping and criterion.• Average 25 days
• Example Criterion• Hit criteria: ≥60 degree turn angle within 2 seconds of initiation• 5 consecutive days of ≥75% performance• 250-300 trials per day
Lever Task – Objective
1 subject, 1 session data from Hulsey et al., 2016
Lever Press Task – Training & Testing
• Shaping• Reward any interaction with the lever
• Training• Adaptive press angle threshold• Adaptive multiple-press hit window• Programmable audible clicks• Average 3 weeks
• Example Criterion• Hit criteria: two presses within 0.5 seconds of initiation• 5 consecutive days of ≥80% performance• 250-300 trials per day
Summary
• Automated forelimb assessments can increase your subject throughput, statistical power, and replicability.
• Vulintus’ MotoTrak System can currently run these automated assessments:
Isometric PullSupinationLever Press…and more in development
• Contact us to talk about tasks you’d like to see automated!
Thank you to our event sponsor
Vulintus' MotoTrak is a complete, modular system designed for computer-supervised training and testing of forelimb movements in rodent models.
Learn more here >
Overcoming Barriers to Translation from Preclinical Research
Copyright 2016 S. Hays, Vulintus and InsideScientific. All Rights Reserved.
Seth Hays, PhDAssistant Professor of Bioengineering, UT Dallas, Director of Preclinical Research, TxBDC
Neural plasticity could be used to treat many neurological diseases if
we could direct it appropriately
• Stroke is a major health issue and a leading cause of disability (Go, 2014)
• Approximately 825,000 cases each year
• Millions of people living with stroke-related disability
• Current rehabilitative therapies are not consistently effective
UF Health
Epidemiology of stroke
Brain changes support recovery after neurological injury
Adapted from Sunderland, 1992
• Rehabilitative training drives adaptive plasticity that is associated with functional recovery (Jones, 2009; Nudo, 1996)
Brain changes support recovery after neurological injury
Adapted from Sunderland, 1992
• Rehabilitative training drives adaptive plasticity that is associated with functional recovery (Jones, 2009; Nudo, 1996)
Brain changes support recovery after neurological injury
Adapted from Sunderland, 1992
• Rehabilitative training drives adaptive plasticity that is associated with functional recovery (Jones, 2009; Nudo, 1996)
Brain changes support recovery after neurological injury
Adapted from Sunderland, 1992
• Rehabilitative training drives adaptive plasticity that is associated with functional recovery (Jones, 2009; Nudo, 1996)
• Methods that enhance plasticity may boost the effects of rehabilitation to improve
VNS paired with forelimb training enhances reorganization of motor cortex
Porter et al., 2011Hulsey et al., 2016
32% increase in distal
159% increase in proximal
Porter et al., 2011Hulsey et al., 2016
32% increase in distal
159% increase in proximal
VNS paired with forelimb training enhances reorganization of motor cortex
Can VNS paired with rehabilitative training enhance recovery after stroke?
Rehab
VNS + Rehab
Experimental Design
Hays et al., NeuroReport, 2014
VNS paired with rehabilitative training improves recovery after ischemic stroke
Khodaparast et al., Neurobiol Dis, 2012
Khodaparast et al., Neurorehab Neural
Repair, 2013
Hays et al., Neuroreport, 2014
Translation of stroke therapy is extremely challenging…
Science Translational Medicine, 2012
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes• Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes• Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
The interaction of age, stroke, and VNS
• Advanced age is a major risk factor for stroke and is associated with worse outcomes
• Plasticity declines with advancing age (Pascual-Leone et al., 2011)
Does advanced age interfere with VNS-dependent enhancement of recovery after stroke?
Hays et al.,
Neurobiol of Aging,
2016
VNS improves recovery of forelimb strength after ischemic stroke in aged rats
Hays et al., Neurobiolof Aging, 2016
VNS improves recovery of forelimb strength after ischemic stroke in aged rats
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Can VNS improve recovery when initiated long after stroke?
• Most interventions become less effective with increasing time after stroke (Biernaskie, 2003; O’Bryant, 2014)
Murphy, 2009
Can VNS improve recovery when initiated long after stroke?
• Most interventions become less effective with increasing time after stroke (Biernaskie, 2003; O’Bryant, 2014)
• An estimated 4 million stroke survivors are left with permanent neurological disability (Tsai, 2011)
VNS paired with rehabilitative training improves recovery after chronic stroke
Khodaparast et al., Neurorehab Neural Repair, 2015
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
Overcoming barriers to translation
Eliminate assessment bias
Ensure sufficient statistical power
Test in models with complicating factors• Hypertension• Diabetes Advanced Age
Determine therapeutic window
Evaluate generalization of benefits
Examine recovery in other models
Fisher et al., Stroke, 2009
• VNS is safe and FDA approved (George et al., 2000)
• 70,000 patients have VNS implants (Englot et al., 2011)
VNS therapy for stroke
• Paired VNS uses less than 1% of the total daily charge
National Clinical Trial NCT01669161
Dawson et al., Stroke, 2015
Early results in stroke patients are encouraging…
VNS + Rehab
Rehab
Undergraduate Researchers:
Maritza Pantoja
Meera Iyengar
Xavier Carrier
Priyanka Das
Iqra Qureshi
Sabiha Sultana
Igor Kushner
Nick Jones
Eric Meyers
Andi Ruiz
David Pruitt
Dr. Navid Khodaparast
Dr. Andrew Sloan
Virginia Land
Sandra Field
Elizabeth Nutting
Suna Burghul
Philip O’Donnell
Reema Patel
Bryan Nguyen
Monisha Thomas
Acknowledgements
FUNDING
SOURCES:
Dr. Patrick Ganzer
Daniel Hulsey
Michael Darrow
Dr. Michael Kilgard
Dr. Robert Rennaker
Thank You!If you have questions for the presenters please contact them by email.
For additional information on the solutions presented in this webinar please visit:
www.vulintus.com
Seth Hays, PhDseth.hays@utdallas.edu
Drew Sloan, PhDdrew@vulintus.com
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