Gait analysis of C57BL/6 mice with complete Freund's adjuvant-induced arthritis using the CatWalk system methods
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Drug treatment
Mice were treated intraperitoneally with indomethacin 0.1 (n = 8), 1 (n = 9) and 10 (n = 9) mg/kg (Sigma-Aldrich, St Louis, MO, USA) or its vehicle (peanut oil, Sigma-Aldrich, USA, n = 10), 48 hours after CFA inoculation based on the gait analysis results (see Results section). The mice were then made to cross the Catwalk walkway at 1, 2 and 3 h post-indomethacin treatment.
Gait analysis
Gait parameters of freely moving mice were measured using the Catwalk gait analysis system (Noldus Information Technology, The Netherlands) as described previously [, ]. Briefly, the CatWalk instrument consists of an enclosed walkway with a glass plate, a fluorescent lamp which emits light inside the glass plate, a high speed colour camera, and a recording and analysis software to assess gait of rodents. Each mouse was placed individually in the CatWalk walkway and allowed to walk freely and traverse from one side to the other of the walkway glass plate. Mice were trained as described previously [ ] and gait parameters assessed before (baseline) and after (daily for 7 days) CFA or PBS inoculation. The recordings were carried out when the room was completely dark, except for the light from the computer screen. Where the mouse paws made contact with the glass plate, light was reflected...
Gait analysis
IV. Stance phase: duration in seconds of contact of a paw with the glass plate in a step cycle i.e. stance phase + swing phase,
Gait analysis
V. Swing phase: duration in seconds of no contact of a paw with the glass plate in a step cycle,
Gait analysis
VI. Duty cycle: stance duration as a percentage of the step cycle duration i.e. stance phase/(stance + swing phases) × 100
Induction of arthritis
Monoarthritis was induced as described previously [ ]. Briefly, mice were anaesthetized with isoflurane and 20 µl of complete Freund's adjuvant (CFA, each ml contained 1 mg of Mycobacterium tuberculosis H37Ra, ATCC 25177, heat killed and dried, 0.85 ml mineral oil and 0.15 ml mannide monooleate) (Sigma-Aldrich, St Louis, MO, USA), was injected intra-articularly into the right hind limb ankle joint through the Achilles tendon using a 29½-gauge needle. Phosphate buffered saline (PBS, 20 µl), was administered in the same way to the control group to evaluate the effects of the injection and volume of liquid injected intra-articulary, because CFA has no vehicle to use as a control.
Methods
Female C57BL/6 mice (8 to 12 weeks old; 20 - 30 g, n = 55) were used and were supplied by the breeding unit at the Health Sciences Center, Kuwait University, Kuwait, and were kept in temperature controlled (24 ± 1°C) rooms with food and water ad libitum. All experiments were performed during the same period of the day (8:00 AM to 4:00 PM) to exclude diurnal variations in pharmacological effects. The animals were handled in compliance with Animal Resources Centre of the Kuwait University Health Sciences Center guidelines and the European Communities Council Directive 86/609 for the care of laboratory animals and ethical guidelines for research in experimental pain with conscious animals [ ]. All procedures were approved by the Animal Resources Centre of the Kuwait University Health Sciences Center.
Gait analysis
VIII. Swing speed: speed of a paw during swing phase i.e. stride length/swing phase duration
Measurement outputs
What raw and processed outputs should exist?
Gait recovery and locomotor function
- Raw artifact
- Per-run gait capture with paw placement, timing, and stride features for each animal
- Processed artifact
- Cleaned gait metrics table and recovery trend summary across timepoints
- Reported as
- Group comparisons of gait indices, stride metrics, or recovery curves
Analysis plan
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Acquisition
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inferred from protocolPreprocessing / cleaning
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inferred from protocolScoring or quantification
Quantify the primary readouts for this experiment: Gait recovery and locomotor function.
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inferred from protocolReporting output
Report representative outputs alongside summary comparisons for Gait recovery and locomotor function.
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Evidence quotes (8)
Mice were treated intraperitoneally with indomethacin 0.1 (n = 8), 1 (n = 9) and 10 (n = 9) mg/kg (Sigma-Aldrich, St Louis, MO, USA) or its vehicle (peanut oil, Sigma-Aldrich, USA, n = 10), 48 hours after CFA inoculation based on the gait analysis results (see Results section). The mice were then made to cross the Catwalk walkway at 1, 2 and 3 h post-indomethacin treatment.
Gait parameters of freely moving mice were measured using the Catwalk gait analysis system (Noldus Information Technology, The Netherlands) as described previously [, ]. Briefly, the CatWalk instrument consists of an enclosed walkway with a glass plate, a fluorescent lamp which emits light inside the glass plate, a high speed colour camera, and a recording and analysis software to assess gait of rodents. Each mouse was placed individually in the CatWalk walkway and allowed to walk freely and traverse from one side to the other of the walkway glass plate. Mice were trained as described previously [ ] and gait parameters assessed before (baseline) and after (daily for 7 days) CFA or PBS inoculation. The recordings were carried out when the room was completely dark, except for the light from the computer screen. Where the mouse paws made contact with the glass plate, light was reflected down and the illuminated contact areas recorded with a high speed colour video camera that was positioned underneath the glass plate connected to a computer that runs the CatWalk software 7.1. The software automatically labelled all the areas containing pixels above the set threshold (7 pixels). Th...
IV. Stance phase: duration in seconds of contact of a paw with the glass plate in a step cycle i.e. stance phase + swing phase,
V. Swing phase: duration in seconds of no contact of a paw with the glass plate in a step cycle,
VI. Duty cycle: stance duration as a percentage of the step cycle duration i.e. stance phase/(stance + swing phases) × 100
Monoarthritis was induced as described previously [ ]. Briefly, mice were anaesthetized with isoflurane and 20 µl of complete Freund's adjuvant (CFA, each ml contained 1 mg of Mycobacterium tuberculosis H37Ra, ATCC 25177, heat killed and dried, 0.85 ml mineral oil and 0.15 ml mannide monooleate) (Sigma-Aldrich, St Louis, MO, USA), was injected intra-articularly into the right hind limb ankle joint through the Achilles tendon using a 29½-gauge needle. Phosphate buffered saline (PBS, 20 µl), was administered in the same way to the control group to evaluate the effects of the injection and volume of liquid injected intra-articulary, because CFA has no vehicle to use as a control.
Female C57BL/6 mice (8 to 12 weeks old; 20 - 30 g, n = 55) were used and were supplied by the breeding unit at the Health Sciences Center, Kuwait University, Kuwait, and were kept in temperature controlled (24 ± 1°C) rooms with food and water ad libitum. All experiments were performed during the same period of the day (8:00 AM to 4:00 PM) to exclude diurnal variations in pharmacological effects. The animals were handled in compliance with Animal Resources Centre of the Kuwait University Health Sciences Center guidelines and the European Communities Council Directive 86/609 for the care of laboratory animals and ethical guidelines for research in experimental pain with conscious animals [ ]. All procedures were approved by the Animal Resources Centre of the Kuwait University Health Sciences Center.
VIII. Swing speed: speed of a paw during swing phase i.e. stride length/swing phase duration
Machine-readable layer
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