Gait analysis of C57BL/6 mice with complete Freund's adjuvant-induced arthritis using the CatWalk system methods
Aim. Evidence-backed execution summary for Gait analysis of C57BL/6 mice with complete Freund's adjuvant-induced arthritis using the CatWalk system methods from Gait analysis of C57BL/6 mice with complete Freund's adjuvant-induced arthritis using the CatWalk system.
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mouse
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Drug treatment
reagent used in the protocol.
- Use
- 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...
Effects of treatment with indomethacin on gait parameters of mice with CFA-induced monoarthritis
reagent used in the protocol.
- Use
- The RH/LH ratios of the static parameters paw pressure and print area were significantly reduced at 48 h after CFA inoculation (2 dpi), similar to the data above (p < 0.01; Figure ). Treatment of monoarthritic mice with indomethacin 10 mg/kg, but not 1 mg/kg, alleviated the paw pressure and print area RH/LH ratio de...
Effects of treatment with indomethacin on gait parameters of mice with CFA-induced monoarthritis
reagent used in the protocol.
- Use
- Effects of indomethacin 1 and 10 mg/kg on (A) weight bearing (measured as ratio of light intensity between right hind (RH) and left hind (LH) limbs and (B) ratio of RH/LH print area of mice with complete Freund's adjuvant (CFA)-induced arthritis. The drugs or their vehicles were administered at 48 hours post-C...
Effects of treatment with indomethacin on gait parameters of mice with CFA-induced monoarthritis
reagent used in the protocol.
- Use
- The RH/LH ratios of the dynamic parameters (stance phase duration, swing phase duration and speed, and duty cycle) were significantly changed at 48 h after CFA inoculation i.e. stance phase duration, swing speed, and duty cycle decreased, whereas swing phase duration increased (p < 0.01; Figure ). Treatment of monoa...
Effects of treatment with indomethacin on gait parameters of mice with CFA-induced monoarthritis
reagent used in the protocol.
- Use
- Effects of indomethacin 1 and 10 mg/kg on (A) ratio of stance phase duration between right hind (RH) and left hind (LH) limbs, (B) ratio of RH/LH swing phase duration, (C) ratio of RH/LH duty cycle {stance phase duration/(stance + swing phases duration) x 100}, and (D) ratio of RH/LH swing speed duration of mice wit...
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...
- Use
- 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...
Acknowledgements
We are grateful to the Department of Pharmacology & Therapeutics, Faculty of Pharmacy, Kuwait University for the CatWalk gait analysis system and to the staff from the Animal Resources Centre, HSC, Kuwait University for their support.
- Use
- We are grateful to the Department of Pharmacology & Therapeutics, Faculty of Pharmacy, Kuwait University for the CatWalk gait analysis system and to the staff from the Animal Resources Centre, HSC, Kuwait University for their support.
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...
- Use
- 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...
Methods
The CatWalk system was used to study gait of freely moving mice with CFA-induced monoarthritis and evaluate pharmacological pain relief in this model of arthritis. CFA (20 µl) was injected intra-articularly into the right hind (RH) limb ankle joint through the Achilles tendon of C57BL/6 mice.
- Use
- The CatWalk system was used to study gait of freely moving mice with CFA-induced monoarthritis and evaluate pharmacological pain relief in this model of arthritis. CFA (20 µl) was injected intra-articularly into the right hind (RH) limb ankle joint through the Achilles tendon of C57BL/6 mice.
Abbreviations
ANOVA: Analysis of variance; CFA: Complete Freund's adjuvant; CNS: Central nervous system; i.p: Intraperitoneally; LH: Left hind limb; LPS: Lipopolysaccharide; PBS: Phosphate buffered saline; RH: Right hind limb; S.E.M: Error of the mean.
- Use
- ANOVA: Analysis of variance; CFA: Complete Freund's adjuvant; CNS: Central nervous system; i.p: Intraperitoneally; LH: Left hind limb; LPS: Lipopolysaccharide; PBS: Phosphate buffered saline; RH: Right hind limb; S.E.M: Error of the mean.
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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?
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-Aldr...
- 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
Statistical analyses were performed using repeated measures two-way analysis of variance (ANOVA) followed by Bonferroni post-tests or one-way ANOVA followed by Dunnett's multipl...
- 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
We are grateful to the Department of Pharmacology & Therapeutics, Faculty of Pharmacy, Kuwait University for the CatWalk gait analysis system and to the staff from the Animal Re...
- 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
Gait parameters of freely moving mice were measured using the Catwalk gait analysis system (Noldus Information Technology, The Netherlands) as described previously [, ]. Briefl...
- 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
How should the outputs become interpretable results?
Acquisition
Capture run-level gait data for each animal and preserve the timepoint or treatment labeling.
inferred from protocolPreprocessing / cleaning
Statistical analyses were performed using repeated measures two-way analysis of variance (ANOVA) followed by Bonferroni post-tests or one-way ANOVA followed by Dunnett's multiple comparison test.
from paperScoring or quantification
Quantify the primary readouts for this experiment: 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-Aldr...; Statistical analyses were performed using repeated measures two-way analysis of variance (ANOVA) followed by Bonferroni post-tests or one-way ANOVA followed by Dunnett's multipl...; We are grateful to the Department of Pharmacology & Therapeutics, Faculty of Pharmacy, Kuwait University for the CatWalk gait analysis system and to the staff from the Animal Re...; Gait parameters of freely moving mice were measured using the Catwalk gait analysis system (Noldus Information Technology, The Netherlands) as described previously [, ]. Briefl....
from paperStatistical comparison
Statistical analyses were performed using repeated measures two-way analysis of variance (ANOVA) followed by Bonferroni post-tests or one-way ANOVA followed by Dunnett's multipl...; Time course of static gait parameters in mice with (A, B, E and F) phosphate buffered saline (PBS) or (C, D, G and H) complete Freund's adjuvant (CFA)-injected intra-artic...; Time course of dynamic gait parameters (A-J) in mice with phosphate buffered saline (PBS) or (K-T) complete Freund's adjuvant (CFA)-injected intra-articularly in the right...; The RH/LH ratios of the static parameters paw pressure and print area were significantly reduced at 48 h after CFA inoculation (2 dpi), similar to the data above (p < 0.01; Figu...
from paperReporting output
Report representative outputs alongside summary comparisons for 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-Aldr..., Statistical analyses were performed using repeated measures two-way analysis of variance (ANOVA) followed by Bonferroni post-tests or one-way ANOVA followed by Dunnett's multipl..., We are grateful to the Department of Pharmacology & Therapeutics, Faculty of Pharmacy, Kuwait University for the CatWalk gait analysis system and to the staff from the Animal Re..., Gait parameters of freely moving mice were measured using the Catwalk gait analysis system (Noldus Information Technology, The Netherlands) as described previously [, ]. Briefl....
inferred from protocolStructured statistical methods
Statistical analyses were performed using repeated measures two-way analysis of variance (ANOVA) followed by Bonferroni post-tests or one-way ANOVA followed by Dunnett's multipl...; Time course of static gait parameters in mice with (A, B, E and F) phosphate buffered saline (PBS) or (C, D, G and H) complete Freund's adjuvant (CFA)-injected intra-artic...; Time course of dynamic gait parameters (A-J) in mice with phosphate buffered saline (PBS) or (K-T) complete Freund's adjuvant (CFA)-injected intra-articularly in the right...; The RH/LH ratios of the static parameters paw pressure and print area were significantly reduced at 48 h after CFA inoculation (2 dpi), similar to the data above (p < 0.01; Figu...
source structuredSource and audit
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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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