Automated Quantitative Measures of Forelimb Function in Rats and Mice

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.

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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

Automated Forelimb Assessments

Common 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


Supination Methods Paper

Isometric Pull Methods Paper

Lever Press Methods Paper

http://www.utdallas.edu/tnpl/files/Meyers_JNeurosciMeth_2016.pdf

http://www.ncbi.nlm.nih.gov/pubmed/23183016


MotoTrak

A modular system for assessing forelimb function

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 Walkthrough

Isometric Pull Task – Setup

Isometric Pull Task – Objective

1 subject, 1 session data from Sloan et al., 2015


Courtesy of Rennaker Lab, UT-Dallas

Isometric Pull Task – Mouse Models

• Modified Setup• Adjusted task dimensions• Smaller manipulandum• More sensitive force sensor

• Liquid reward vs. pellet reward

Courtesy of April Becker, Goldberg Lab, UTSW Medical Center

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 – Setup

Supination Task – Objective

1 subject, 1 session data from Meyers et al., 2016


Courtesy of Eric Meyers, Rennaker Lab, UT-Dallas

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 – Setup

Lever Task – Objective

1 subject, 1 session data from Hulsey et al., 2016


Courtesy of Hays Lab, UT-Dallas

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!

mailto:[email protected]?subject=MotoTrak%20Webinar%20(InsideScientific)

mailto:[email protected]?subject=MotoTrak%20Webinar%20(InsideScientific)

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 >

http://www.vulintus.com/mototrak/








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)










• Methods that enhance plasticity may boost the effects of rehabilitation to improve

Neuromodulator Release

Neural Activity

Plasticity

+

Insert title…

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?

Neuromodulator Release

Rehabilitation

Recovery

+

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




Test in models with complicating factors• Hypertension• Diabetes• Advanced Age





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

http://www.sciencedirect.com/science/article/pii/S0197458016300227

Hays et al., Neurobiolof Aging, 2016

VNS improves recovery of forelimb strength after ischemic stroke in aged rats




Test in models with complicating factors• Hypertension• Diabetes Advanced Age













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)

Khodaparast et al., Neurorehab Neural Repair, 2015


VNS paired with rehabilitative training improves recovery after chronic stroke

Khodaparast et al., Neurorehab Neural Repair, 2015


















Does VNS-dependent recovery generalize to similar forelimb tasks?

Enhancement of forelimb recovery generalizes to similar motor tasks

Enhancement of forelimb recovery generalizes to similar motor tasks

















Assessing VNS paired with rehab to improve recovery after peripheral nerve injury

VNS may improve recovery after peripheral nerve injury









• 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

Clinical translation of VNS therapy…

National Clinical Trial NCT01669161

Dawson et al., Stroke, 2015

Early results in stroke patients are encouraging…

VNS + Rehab

Rehab

http://stroke.ahajournals.org/content/early/2015/12/08/STROKEAHA.115.010477.full.pdf+html

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, [email protected]

Drew Sloan, [email protected]




http://www.vulintus.com/

http://www.vulintus.com/