Using Touchscreen Operant Systems to Study Cognitive Behaviors in Rodents

Experts present methodology and best practices of Bussey-Saksida Touchscreen Systems used for the cognitive evaluation of rodents.

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An Introduction to Touchscreen Systems

Our Talk Today

1. Background

2. Hardware Requirements

3. ABET II Touch Software

4. Validated Tasks

5. Integrating with other systems

• The Cambridge Neuropsychological Test Automated Battery (CANTAB) was first developed 30 Years Ago

• Designed from a knowledge animal behavioral tests

• Proven tests sensitive to specific brain areas

• Presented on touchscreens

• Abstract shapes used to eliminate differences in language and culture.

Touchscreen Testing Background – Human CANTAB Tests

• Some of the Human tasks made suitable for Non-Human Primates

• Complex tasks broken down into their constituent elements

• Tests proven to be sensitive to the same specific brain areas as the human tests

Touchscreen Testing Background – Monkey CANTAB

• Work gone full circle to develop equivalent tasks for rodent

• Complex tasks broken down into their constituent elements

• Tests proven to be sensitive to the same specific brain areas as the human tests

• Initial work in standard operant chambers with touchscreen bolted

Rodent Touchscreen Development

• Chamber design – Keep the animal focused on the task

• Effective training

• Flexible data analysis

• Validated Tasks – Functional equivalence to Human Tasks

• On going development of tasks

Important Considerations in designing translational Rodent Tasks

• use in place of levers – e.g. PR task

• use in place of nose pokes – e.g. 5-choice serial reaction time task

• Response to different images adds further function over more traditional operant chambers

• A wide range of stimuli positions available

• Further flexibility by adding elements from more standard chambers – levers, lights, sound etc.

• Addition of video adding a further element to testing.

Touchscreens Flexibility

Additional information can be gained by seamlessly integrating with other cognitive measures:

Electrophysiology Recording

Video Tracking

Optogenetics

Touchscreen Integration…

Hardware & Software

Hardware Requirements

System Layout

1. Single Computer

2. Digital I/O Interface

3. Four TouchscreenChambers

Computer

Box 1 Box 2 Box 3 Box 4

Digital I/O Interface

Video & Touch Data

Inputs/Outputs

Mouse Chamber

1. Touchscreen

2. Mask (paradigm specific)

3. Reward Pump

4. Reward Bottles

5. Perforated Floor

6. Waste Pan

7. Ambulatory IR Beams

1

2

3

45

6

7

Mouse Chamber

1. Touchscreen

2. Trapezoidal Walls & Lid

3. Reward Trough

4. Reward Bottles

5. Ambulatory IR Beams

6. Pull-out roller shelf

7. House Light/Fan/Speaker

1

2

3

4

5

6

7

Touchscreens – IR Technology

Lots of ‘touch’ technologies available…IR Technology provides:

No force requirements

Behaviorally similar to nosepoke

No interference with telemetry

Easy identification of ‘discrete’ and ‘intended’ touches

** Other touch technologies can be used, but not as effectively.

• Modular Walls – add standard operant accessories (levers, lights, nosepokes, ect.)

• Tether compatible walls, lid, and reward trough – for canulas, neural recording, or optogentics fibers

• Easy-Install Kit – four chambers neatly mounted on their own car for mobility and ease of use

• Grid Shock – Aversive foot shock

• TTL Breakout box – Trigger 3rd party equipment

Additional Options

Touchscreens vs. Operant

• Versatile – can run multiple paradigms on a single chamber

• Flexible – change spatial location on a trail by trial basis automatically

• Powerful – use custom images, control stimuli locations, adjust the paradigm

ABET II TOUCHAnimal Behavioral Environment Test

ABET II Touch

Tab Name Description

Environment Designer Identify what hardware (lights, lever, reward pump) is connected

Schedule Designer Create, open, and edit schedule files which control the chambers

Execution Manager Run schedule files in the chambers with independent control

Analysis Set DesignerCreate, open, and edit analysis files which pull out important metrics from the RAW data

Data ViewerReview RAW data or run analysis files to create daily performance reports

ABET II Touch – Environment Designer

1. Chambers

2. Touch monitor settings

3. Inputs – IR Beams, Levers, NosePokes

4. Outputs –HouseLight, Reward, Tone, TrayLight

1

3 4

2

Identify what hardware (lights, lever, reward pump) is connected

1

3

4

2

ABET II Touch – Schedule Designer

Create, open, and edit schedule files which control the chambers

1. Schedule information

2. Variables, Lists, Arrays

3. Schedule structure and progression

4. Code view of selected group

13

4

2

ABET II Touch – Execution Manager

Run schedule files in the chambers with independent control

1. Chambers

2. Session end criterion – Max trials or time

3. Database selection for data saving

4. Session Information – Animal ID, user, weight, dose, …

ABET II Touch – Analysis Set Designer

Create, open, and edit analysis files which pull out important metrics from the RAW data

1

3

4

1. Analysis file information

2. Analysis Item – Path, counter, evaluation

3. Analysis Point –Start/stop, variable, time point

4. Code view of selection

2

13

4

2

ABET II Touch – Data Viewer

Review RAW data or run analysis files to create daily performance reports

1. Database search for specific sessions

2. Analysis tab to run analysis files

3. Various views of group statistics

4. Report Editor –create your own spreadsheet design and easily export

ABET II Touch - Features

Setup and control various hardware configurations

Easily create and modify your schedules

Load your own images into the schedules

Independent control and millisecond timing

Quickly view “meaningful data” after running

Guided interface reduces learning curve

Additional Online Resources

Download the Bussey-Saksida Rodent Touch System Brochure

Join our Rodent Touchscreen Users Forum

CHAMBERS AND COMPONENTS FOR RATS

CHAMBERS FOR ELECTROPHYSIOLOGY

CHAMBERS AND COMPONENTS FOR MICE

Validated Tasks

Validated Tasks – A Turnkey Solution

1. The Tasks

2. Application to different Neural Area

3. Training Times

4. Rodent strains

5. The PAL task

• Pairwise / Visual Discrimination (PD)

• Pairwise Associate Learning (PAL)

• Visuomotor Conditional Learning (VMCL)

• Location Discrimination (LD)

• Trial Unique Nonmatching to Location (TUNL)

• 5-Choice Serial Reaction Time Task (5-CSRTT)

• Autoshaping (AUTO)

• Extinction (EXT)

• Progressive Ratio (PR); Effort Related Choice (ERC)

• Rodent Continuous Performance Test (rCPT)

The Tasks

TaskExample Neural Systems Involved

Clinical Area Showing Impairment

Pairwise / Visual Discrimination (PD)

Prefrontal Cortex, Perirhinal Cortex, Striatum, Dopamine System, Cholinergic System, NMDA receptors

Huntingdon’s, Schizophrenia, Parkinson’s

Pairwise Associate Learning (PAL)

Hippocampus, Cholinergic System, NMDA Receptors, AMPA Receptors

Alzheimer’s, Schizophrenia

Visuomotor Conditional Learning (VMCL)

Dorsal Striatum, Posterior Cingulate Cortex Huntingdon’s, Parkinson’s

Location Discrimination (LD) Hippocampus, NeurogenesisAlzheimer’s, Schizophrenia, Depression

Trial Unique Nonmatching to Location (TUNL)

Hippocampus, Cholinergic System, NMDA Receptors Alzheimer’s

TaskExample Neural Systems Involved

Clinical Area Showing Impairment

5-Choice Serial Reaction Time Task (5-CSRTT)

Prefrontal Cortex, Basal Forebrain, Serotonin (Impulsivity), Cholinergic System (Accuracy), Noradrenaline (Distraction), Dopamine (Motivation)

Huntingdon’s, Schizophrenia, Alzheimer’s, Depression, ADHD, OCD

Autoshaping (AUTO)Ventral Striatum, Amygdala Anterior Cingulate Cortex

Huntingdon’s

Extinction (EXT) Infralimbic, Ventral Striatum, Amygdala ADHD, OCD

Progressive Ratio (PR); Effort Related Choice (ERC)

Dopamine systemHuntingdon’s, Schizophrenia, Alzheimer’s

Rodent Continuous Performance Test (rCPT)

Hippocampus, Cholinergic System, NMDA Receptors

Schizophrenia, Alzheimer’s, ADHD, OCD

Task Training Time - Rats Training Time - Mice

Pre-training to touch image and initiate (PD, PAL, LD, VMCL, TUNL)

1-2 weeks1-2 weeks (e.g. 7-8 week old C57BL6J mice: 5 days)

Pairwise/Visual Discrimination (PD) 5-7days 5-7 days for young mice

Pairwise Associate Learning (PAL) 35-45 sessions to 80% 35-45 sessions to 70%

Visuomotor Conditional Learning Approximately 20 sessions Approximately 20 sessions

Location Discrimination (LD) 2-4 weeks 2-4 weeks

Trial Unique Nonmatching to Location (TUNL)

Approximately 4 weeks 14-16 sessions

5-Choice Serial Reaction Time Task(5-CSRTT)

~30 sessionsPre-training (av. 10 days) and 3 weeks to 80% @ 2sec

Task Training Time - Rats Training Time - Mice

Autoshaping (AUTO) Several sessions Several sessions

Extinction (EXT)~4 days training and a few days extinction

~4 days training and a few days extinction

Progressive Ratio (PR); Effort Related Choice (ERC)

16 days to train mice from the first habituation session to reach stable PR performance.

Rodent Continuous Performance Test (rCPT)

~ 24 sessions

Rats

• Lister hooded, Pigmented DA strain, Sprag-Dawley, Long-Evans

• Genetic Absence Epilepsy Rats from Strasbourg (GAERS) -showed a significant deficit on PD, compared to the non-epileptic control (NEC) strain. (John Howland – work to be published)

Rodent strains used with Touchscreen chambers

Mice

• C57BL/6, C57BL/6J, DBA/2J, C57BL/6JRj, Heterozygous NR2A (HET), 5HTT null KO and HET, M1R-/- (show increased perseveration), TgCRND8, DLg1, Dlg2, Dlg3 & Dlg4 mutations

• Showing deficits in the PD task - R6/2, NR2A KO

• Showing impairment in the 5-CSRTT –3xTgAD, M1R-/- (show increased perseveration)

Rodent strains used with Touchscreen chambers

Translational Tasks

Paired associate learning

5-Choice serial reaction time

Spatial Working Memory

Frontal Temporal

Frontal

Frontal Parietal

Set shifting

Visual Discrimination and reversal

Continuous Performance Task Frontal Temporal

Frontal

Frontal

Example Patient Profile

-2.5

-2

-1.5

-1

-0.5

0

Pattern Spatial PAL IDED

Alzheimer’s

Parkinson’s

Standard deviations

fromcontrolmean

Paired Associative Learning (PAL)

Highly sensitive to cognitive decline in Alzheimer’s disease

Activates neural correlates for new learning & memory

Paired Associative Learning (PAL) – Human Task

1. Boxes open in a random order to reveal an abstract shape

2. Then the shapes are shown in a random order in the centre of the screen. The subject touches the boxes where they first saw the shape

3. The process is repeated until no mistakes are made

0

20

40

60

80

100

120

140

PA

L e

rro

rs

-4 0 4 8 12 16 20 24 28

Time point (months)

pDAT

QD1

QD2

Normal controls

Fowler, K. S., Saling, M. M., Conway, E. L., Semple, J. M. and Louis, W. J. (1997) Computerized neuropsychological tests in the early detection of dementia: prospective findings. Journal of the International Neuropsychological Society, 3, 139-146.

Sensitivity of PAL to decline in Alzheimer’s Disease

Sensitivity of PAL to decline in Alzheimer’s Disease

• The 34 error cut-off (shown by red line) would have discriminated at the 6-month point who in the QD group would go on to receive AD diagnoses and who would not.

Sensitivity of PAL to decline in Alzheimer’s Disease

Swainson, R., Hodges, J. R., Galton, C. J., Semple, J., Michael, A., Dunn, B. D., Iddon, J. L., Robbins, T. W. and Sahakian, B. J. (2001) Early detection and differential diagnosis of Alzheimer's disease and depression with neuropsychological tasks. Dementia and Geriatric Cognitive Disorders, 12 (4), 265-280.

Monkey PAL

Monkey PAL

Monkey PAL

• Rhesus Monkeys readily acquire the CANTAB-PAL task and performance is graded by task difficulty

• Repeated attempts to learn a given set of stimulus-location associations improves completion accuracy

• Ketamine and scopolamine both impair the initial attempt in a dose-dependent fashion.

• Monkeys treated with ketamine require additional attempts to learn a set of stimulus-location associations

• Monkeys treated with scopolamine make fewer attempts and do not improve with repeated opportunities

• The CANTAB-PAL task may serve to distinguish a lack of sustained attention from impaired memory

Taffe et al, Psychopharmacology (2002), 160, 253-262

Ro

den

t PA

LS+ S-

S- S+

S- S+

S+ S-

S- S+

S+ S-

Rodent PAL

PAL Training

1. Animal handling

2. Food Deprivation

3. Habituation to touchscreen chamber

4. Pavlovian training to associate reward with images on the screen

5. Training to touch the screen

6. Training to initiate the tasks

7. Training to ignore blank windows

8. The PAL task

PAL Task Manipulations and Validation

S+ S-

sPA

L

Antagonism of NMDA (shown above) or AMPA receptors in the hippocampus impairs performance in rats on dPAL, but not on sPAL

Talpos JC, Winters BD, Dias R, Saksida LM, Bussey TJ, Psychopharmacology 2009 157

S+ S-

dPA

L

PAL Task –a translational task

Humans with DLG2 mutations and Dlg2-knockout mice both show impairment on the dPALtask.

Nithianantharajah et al, Nature Neuroscience, 201316

Tasks overview

• Wide range of validated tasks

• Covering a range of cognitive functions

• Training from a few days to 1-2 months

• Used with a range of rat and mouse strains, including KO strains.

• Offers an alternatives to standard lever and nose-poke responses

Systems Integration

Electrophysiology Integration

Chambers compatible for neural recordings:

Sound Attenuation Chamber lined with EMI resistant paint/stainless steel

Touchscreens have additional EMI reducing film

Electrical components/cables are shielded

TTL event lines to mark events on neural data files

Optogenetics & Video Tracking

Increased height Sound Attenuation Chambers

Additional TTL output lines for triggering/marking events

Walls and reward receptacle have been designed for animals with head caps

Upper rolling shelf for easy mounting

• All the features of previous electrophysiology chambers

• Integrated auto-retract tether with shielded cable

• Digital headstage and range of electrodes

• Synchronous video acquisition and event markers in the neural data

• Each component is designed specifically for use in the touchscreenchambers

Fully Integrated Solution

Tethered and Wireless options

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.lafayetteinstrument.comJulie Gill, PhD

jgill@campdeninstruments.com

Matt Croxallmatt@lafayetteinstrument.com