Sep 20, 2024
Behavioral analyses in mice
- Nicolas Giguère1,2,
- louis-eric.trudeau Trudeau1,2
- 1Department of Pharmacology and Physiology, Faculty of Medicine, Université de Montréal;
- 2Department of Neurosciences, Faculty of Medicine, Université de Montréal
Protocol Citation: Nicolas Giguère, louis-eric.trudeau Trudeau 2024. Behavioral analyses in mice . protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzj234lx1/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 02, 2024
Last Modified: September 20, 2024
Protocol Integer ID: 104926
Keywords: ASAPCRN, different behavioural assay, mice, nose poking, grip strength test, test, learning task
Funders Acknowledgements:
Aligning Science Across Parkinson's (ASAP)
Grant ID: ASAP-000525
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Abstract
This collection of protocols describes different behavioural assays performed in mice including, grip strength test, pole test, rotarod, open field and nose poking learning task.
Guidelines
This protocol needs prior approval by the users' Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee
Materials
- Grip strength apparatus (BioSeb instruments, BIO-GS3, France)
Equipment
Grip strength apparatus (BioSeb instruments, BIO-GS3, France)
NAME
BioSeb instruments
BRAND
BIO-GS3
SKU
LINK
- Rotarod apparatus (Harvard Apparatus, LE8205, USA)
Equipment
rotarod apparatus (Harvard Apparatus, LE8205, USA)
NAME
Rota Rod with Touchscreen, 5 Mice
TYPE
Rota Rod
BRAND
76-0770
SKU
LINK
- Infrared actimeter (Superflex sensor version 4.6, Omnitech) using the Fusion software (v5.6 Superflex Edition, RRID:SCR_017972)
- 0.9% sodium chloride saline solution (Halyard, #cat 116)Halyard Saline Unit Single Dose 0.9% Sodium Chloride Solution - 15mL - Box 24Halyard healthCatalog #116
- Cocaine hydrochloride 20 mg/kg (Medisca, cat# 53-21-4, Canada)Cocaine HydrochlorideMedisca Inc.
- D-amphetamine sulfate at 5 mg/kg (Tocris, 2813, UK)D-Amphetamine sulfateTocrisCatalog #2813
- SCH23390-HCl 50 µg/kg (Sigma, D-054, Canada)R( )-SCH-23390 hydrochlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #D-054
- Quinpirole-HCl 0.2 mg/kg (Sigma, Q-102, Canada)(−)-Quinpirole hydrochlorideMerck MilliporeSigma (Sigma-Aldrich)Catalog #Q-102
- Raclopride L-tartrate 1 mg/kg (Sigma, R-121, Canada)S(−)-Raclopride ( )-tartrate saltMerck MilliporeSigma (Sigma-Aldrich)Catalog #R-121
Troubleshooting
Grip strength test
2h 30m
A grip strength apparatus (BioSeb instruments, BIO-GS3, France) was used to evaluate paw force of 10-12-week-old male or female mice.
Procedure:
Habituate the mouse in the testing room for at least 01:00:00 .
1h
Hold the mouse firmly by the tail and slowly lower it on the grid of the grip strength apparatus until the forepaws grasp the middle of the grid.
Lower the mouse to a horizontal position and slowly pull following the axis of the sensor until the mouse releases its grasp on the grid.
Repeat the test 3 times with 30 min resting period between each trial.
- Repeat the test for 00:30:00 (1/3).
- Repeat the test for 00:30:00 (2/3).
- Repeat the test for 00:30:00 (3/3).
1h 30m
Once testing is completed, place the mouse back into its home cage and clean the apparatus before testing the next mouse.
Pole test
1h
The test was conducted with a homemade 48 cm metal rod of 1 cm diameter covered with adhesive tape to facilitate traction. Male or female mice of 10-12-weeks of age were used.
Procedure:
Habituate the moude in the testing room for at least 01:00:00 .
1h
Hold the mouse on the surface of your hand and bring it up to the top of the pole.
Position the mouse head-up at the top of the pole.
Record the time required to turn (t-turn) and to climb down completely.
Once testing is completed, place the mouse back into its home cage and clean the metal rod before testing the next mouse.
Rotarod
5h 21m
Motor coordination was evaluated with a rotarod apparatus (Harvard Apparatus, LE8205, USA) on 10-12-week-old male or female mice.
Mice were pre-trained on the rotarod for two consecutive days to reach a stable performance and then tested on the third day.
Procedure:
Day 1
- Habituate the mouse in the testing room for at least 01:00:00 .
- Place the mouse on the rotarod at 4 rpm for up to 00:01:00 or a maximum or 3 trials.
- Once testing is completed, place the mouse back into its home cage and clean the apparatus before testing the next mouse.
1h 1m
Day 2
- Habituate the mouse in the testing room for at least 01:00:00 .
- Place the mouse on the rotarod with an accelerated rotation of 4 rpm -40 rpm for 00:10:00 .
- Repeat 2 more times with 30 minutes resting period between each trial.
- Repeat for 00:30:00 (1/2).
- Repeat for 00:30:00 (2/2).
2h 10m
Day 3
- Habituate the mouse in the testing room for at least 01:00:00 .
- Place the mouse on the rotarod with an accelerated rotation of 4 rpm -40 rpm for 00:10:00 .
- Repeat 2 more times with 30 minutes resting period between each trial.
- Repeat for 00:30:00 (1/2).
- Repeat for 00:30:00 (2/2).
- Record the latency to fall of all 3 trials.
2h 10m
Open field
1h
The locomotor behavior of 11-12-week-old male or female mice was recorded using an infrared actimeter (Superflex sensor version 4.6, Omnitech) using the Fusion software (v5.6 Superflex Edition, RRID:SCR_017972). A chamber partition was used to test two mice at the same time. Subjects were not given time to acclimate to the chamber and spent a total of 01:00:00 in the chamber with the following protocol:
1h
Habituate the mouse in the testing room for at least 01:00:00 .
1h
Place the mouse in the chamber and record basal locomotion for 00:20:00 .
20m
After 00:20:00 , pause the recording and administer drugs or saline. In the present study, this was one of the following substances:
- 0.9% sodium chloride saline solution (Halyard, #cat 116)
- Cocaine hydrochloride 20 mg/kg (Medisca, cat# 53-21-4, Canada)
- D-amphetamine sulfate at 5 mg/kg (Tocris, 2813, UK)
- SCH23390-HCl 50 µg/kg (Sigma, D-054, Canada)
- Quinpirole-HCl 0.2 mg/kg (Sigma, Q-102, Canada)
- Raclopride L-tartrate 1 mg/kg (Sigma, R-121, Canada)
20m
Immediately place the mouse back into the chamber and record locomotion for another 00:40:00 .
40m
Once testing is completed, place the mouse back into its home cage and clean the open field boxes before testing the next mouse.
Nose poking operant learning task
30m
Appetitively motivated operant learning was assessed using the open source Feeding Experimentation Device 3 (FED3) (https://doi.org/10.7554/eLife.66173). Individually housed animals were evaluated in a 12h/12h light/dark cycle.
Procedure:
Place the feeding devices in the home cages under a free feeding paradigm (automatically dispenses a new pellet each time the mouse collects the pellet in the well) for 2 days.
After 2 days, change the device mode to a fixed ratio (FR1) paradigm (the mouse learns to nose poke on the active port to receive a single pellet).
Record nose-poking rate and poke efficiency for 3 consecutive days.
After 3 days, change the device mode to a repeating progressive ratio schedule (nose-poking requirement begins at FR1 and increases by 1 poke each time a pellet is earned)
o When mice fail to poke on either active or inactive port for 00:30:00 , the ratio is automatically reset to FR1.
30m
Record nose-poking rate and poke efficiency for 3 consecutive days.
Once testing is completed, remove the feeding device from the home cage and clean it before testing the next session.