Gait Assessment of Pain and Analgesics: Comparison of the DigiGait™ and CatWalk™ Gait Imaging Systems methods
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CatWalk ™ XT Analysis
Before experiments, animals were trained to make uninterrupted runs within 3.0 s. Measurements were taken at 7 days or 9 days after SNI surgery or CFA injection. Each rat was placed on the walkway repeatedly at intervals of at least 5 min to complete three uninterrupted runs for further analyses.
CatWalk ™ XT Analysis
Analyses of these recordings yielded many parameters, of which the following have been the most acknowledged and applied in pain models [ ]: (1) Coordination data (CatWalk-swing and CatWalk-duty cycle). CatWalk-swing is the duration of the swing phase during walking. Since the durations of the stance and swing phases depend on the walking speed and degree of dysfunction, these parameters are transformed to a fraction of the total step duration according to the following formula: CatWalk-duty cycle = (time in stance phase/time in single step). (2) Area data (CatWalk-max contact area and CatWalk-print area). CatWalk-max contact area is the maximum of total floor area in cm 2 from single images of the paw in the stance phase. CatWalk-print area is the total floor area in cm 2 summed from all images of the paw in the stance phase (compared to max contact area, this parameter reflects the...
DigiGait ™ Analysis
Before experiments, animals were trained to make uninterrupted runs for at least 3 step-cycles at a speed of at least 10 cm/s. Measurements were taken at 7 days or 9 days after SNI surgery or CFA injection. Each rat was placed on the treadmill repeatedly at intervals of at least 5 min to complete three uninterrupted runs (containing at least 3 step-cycles per run) for further analyses. The velocity of the treadmill was initially set at 10 cm/s and was gradually increased or decreased in 1 cm/s steps depending upon the rat's performance, to minimize the stress induced by forced movement.
Efficiency of DigiGait ™ and CatWalk ™ in Reflecting the Severity of Neuropathic Pain in the SNI Model
For the coordination data, the time of swing, duration (swing), and the percentage in stance (duty cycle) during a single step-cycle were calculated in both DigiGait ™ and CatWalk ™. On the treadmill or catwalk track, rats initially contacted the floor with the palm, then transferred to the toes, and ended with the heel. Transmitted by the remaining uninjured sural nerve, mechanical hypersensitivity due to central sensitization prompted the SNI rats to avoid bearing weight on the injured left hindpaw, resulting in a longer swing duration of that limb than the uninjured right hindlimb. In representative CatWalk ™ graphs of consecutive prints (Fig. D), the saline-swing (LH/RH) was 2.000, calculated as (19-11)/(19-15), and PGB decreased this ratio to 1.500, calculated as (28-16)/(28-20); the saline-duty cycle (LH/RH) was 0.733 [(11/15)/(15/19)], and PGB normalize...
Efficiency of DigiGait ™ and CatWalk ™ in Reflecting the Severity of Neuropathic Pain in the SNI Model
Furthermore, we analyzed the correlations between mechanical threshold and each gait parameter in DigiGait ™ and CatWalk ™. For the coordination data in both systems, significant correlations were found for the duration of swing ( n = 22, DigiGait ™: P = 0.046; CatWalk ™: P = 0.049) but not for duty cycle ( n = 22, DigiGait ™: P = 0.086; CatWalk ™: P = 0.060) (Fig. A, B). For the area data, DigiGait ™ -projected area and CatWalk ™ -max contact area and print area displayed no correlation with punctate allodynia (Fig. A, B). In addition, no correlation was detected with CatWalk ™ -mean intensity (Fig. G). Taken together, these statistics indicated that coordination data (swing) reflected the mechanical hypersensitivity in SNI rats, but the intensity and area data did not. Fig. 3 Coordination data are pain...
Efficiency of DigiGait ™ and CatWalk ™ in Detecting Alleviation of Gait Abnormalities in Response to Anal...
PGB (Lyrica®, Pfizer), FDA-approved for peripheral and central neuropathic pain, and especially associated with diabetic peripheral neuropathy, postherpetic neuralgia, and spinal cord injury in adults, was administered as an analgesic in the 22 SNI rats [, ]. PGB is an analogue of γ-aminobutyric acid with high affinity and selectivity for the α2δ subunit of voltage-sensitive Ca 2+ channels (~217 amino-acids) and inhibits their activity during pain transduction [ ]. Adverse effects of PGB such as balance disorder (~4%) and peripheral edema (8%) might confuse the observation of changes in gait parameters [ ]. However, the balance disorder is relatively severe and cannot be counteracted by calculations, so it is necessary to determine the proper dose of PGB with moderate analgesic action and minimal adverse effects. The maximal plasma level of PGB occurs 1 hR...
Efficiency of DigiGait ™ and CatWalk ™ in Reflecting the Severity of Inflammatory Pain in the CFA Model
Furthermore, correlation analyses were conducted between mechanical threshold and each gait parameter in DigiGait ™ and CatWalk ™. For the coordination data, in both systems, strong correlations were found with duration of swing ( n = 22, DigiGait ™: P = 0.003; CatWalk ™: P = 0.003) and duty cycle ( n = 22, DigiGait ™: P = 0.002; CatWalk ™: P = 0.130) (Fig. A, B). For area data, (1) DigiGait ™ -projected area and (2) CatWalk ™ -max contact area and print area displayed no correlation with punctate allodynia (Fig. A, B). In addition, a significant correlation was detected with CatWalk ™ -mean intensity ( n = 22, P = 0.050) (Fig. G). Thus, correlation statistics indicated that the coordination and intensity data, but not the area data, were correlated with the mechanical hypersensitivity in CFA rats. Fig....
Efficiency of DigiGait ™ and CatWalk ™ in Detecting Alleviation of Gait Abnormalities in Response to Anal...
A previous study in rats with chronic arthritis showed that 9 mg/kg subcutaneous (s.c.) tramadol slightly reduced the mechanical allodynia, 29 mg/kg s.c. tramadol induced moderate alleviation of punctate allodynia and joint hyperalgesia, and 88 mg/kg s.c. completely rescued these sensory abnormalities to baseline level at 30 min post-administration [ ]. In a sodium monoiodoacetate-induced osteoarthritis model, 100 mg/kg oral tramadol (2 h prior to tests) normalized the CatWalk ™ -max contact area and swing speed [ ]. Taken together, these data led us to select the dose of 30 mg/kg tramadol, and the behavioral assessments were performed during 1 h-2 h after i.p. tramadol injection.
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Gait recovery and locomotor function
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- Per-run gait capture with paw placement, timing, and stride features for each animal
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Quantify the primary readouts for this experiment: Gait recovery and locomotor function.
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Report representative outputs alongside summary comparisons for Gait recovery and locomotor function.
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Evidence quotes (8)
Before experiments, animals were trained to make uninterrupted runs within 3.0 s. Measurements were taken at 7 days or 9 days after SNI surgery or CFA injection. Each rat was placed on the walkway repeatedly at intervals of at least 5 min to complete three uninterrupted runs for further analyses.
Analyses of these recordings yielded many parameters, of which the following have been the most acknowledged and applied in pain models [ ]: (1) Coordination data (CatWalk-swing and CatWalk-duty cycle). CatWalk-swing is the duration of the swing phase during walking. Since the durations of the stance and swing phases depend on the walking speed and degree of dysfunction, these parameters are transformed to a fraction of the total step duration according to the following formula: CatWalk-duty cycle = (time in stance phase/time in single step). (2) Area data (CatWalk-max contact area and CatWalk-print area). CatWalk-max contact area is the maximum of total floor area in cm 2 from single images of the paw in the stance phase. CatWalk-print area is the total floor area in cm 2 summed from all images of the paw in the stance phase (compared to max contact area, this parameter reflects the entire paw area from the third toe to the heel). (3) Intensity data (CatWalk-mean intensity). CatWalk-mean intensity is the mean intensity of the contact area of the hindpaw during a step cycle. This parameter is expressed in arbitrary units.
Before experiments, animals were trained to make uninterrupted runs for at least 3 step-cycles at a speed of at least 10 cm/s. Measurements were taken at 7 days or 9 days after SNI surgery or CFA injection. Each rat was placed on the treadmill repeatedly at intervals of at least 5 min to complete three uninterrupted runs (containing at least 3 step-cycles per run) for further analyses. The velocity of the treadmill was initially set at 10 cm/s and was gradually increased or decreased in 1 cm/s steps depending upon the rat's performance, to minimize the stress induced by forced movement.
For the coordination data, the time of swing, duration (swing), and the percentage in stance (duty cycle) during a single step-cycle were calculated in both DigiGait ™ and CatWalk ™. On the treadmill or catwalk track, rats initially contacted the floor with the palm, then transferred to the toes, and ended with the heel. Transmitted by the remaining uninjured sural nerve, mechanical hypersensitivity due to central sensitization prompted the SNI rats to avoid bearing weight on the injured left hindpaw, resulting in a longer swing duration of that limb than the uninjured right hindlimb. In representative CatWalk ™ graphs of consecutive prints (Fig. D), the saline-swing (LH/RH) was 2.000, calculated as (19-11)/(19-15), and PGB decreased this ratio to 1.500, calculated as (28-16)/(28-20); the saline-duty cycle (LH/RH) was 0.733 [(11/15)/(15/19)], and PGB normalized this ratio to 0.800 [(16/28)/(20/28)].
Furthermore, we analyzed the correlations between mechanical threshold and each gait parameter in DigiGait ™ and CatWalk ™. For the coordination data in both systems, significant correlations were found for the duration of swing ( n = 22, DigiGait ™: P = 0.046; CatWalk ™: P = 0.049) but not for duty cycle ( n = 22, DigiGait ™: P = 0.086; CatWalk ™: P = 0.060) (Fig. A, B). For the area data, DigiGait ™ -projected area and CatWalk ™ -max contact area and print area displayed no correlation with punctate allodynia (Fig. A, B). In addition, no correlation was detected with CatWalk ™ -mean intensity (Fig. G). Taken together, these statistics indicated that coordination data (swing) reflected the mechanical hypersensitivity in SNI rats, but the intensity and area data did not. Fig. 3 Coordination data are pain-related gait parameters in the SNI model. A Correlation analysis between mechanical threshold (x-axis) and DigiGait-swing (left y-axis) and DigiGait-duty cycle (right y-axis). Linear regression, * P < 0.05. B Correlation analysis between mechanical threshold (x-axis) and CatWalk-swing (l...
PGB (Lyrica®, Pfizer), FDA-approved for peripheral and central neuropathic pain, and especially associated with diabetic peripheral neuropathy, postherpetic neuralgia, and spinal cord injury in adults, was administered as an analgesic in the 22 SNI rats [, ]. PGB is an analogue of γ-aminobutyric acid with high affinity and selectivity for the α2δ subunit of voltage-sensitive Ca 2+ channels (~217 amino-acids) and inhibits their activity during pain transduction [ ]. Adverse effects of PGB such as balance disorder (~4%) and peripheral edema (8%) might confuse the observation of changes in gait parameters [ ]. However, the balance disorder is relatively severe and cannot be counteracted by calculations, so it is necessary to determine the proper dose of PGB with moderate analgesic action and minimal adverse effects. The maximal plasma level of PGB occurs 1 h-2 h after i.p. administration [ ]. In addition, the strongest analgesic action and limited side-effects occur during this period. Previous studies of PGB in a partial sciatic nerve ligation model have confirmed a slight (12.5 mg/kg) and moderate (25 mg/kg) alleviation of punctate a...
Furthermore, correlation analyses were conducted between mechanical threshold and each gait parameter in DigiGait ™ and CatWalk ™. For the coordination data, in both systems, strong correlations were found with duration of swing ( n = 22, DigiGait ™: P = 0.003; CatWalk ™: P = 0.003) and duty cycle ( n = 22, DigiGait ™: P = 0.002; CatWalk ™: P = 0.130) (Fig. A, B). For area data, (1) DigiGait ™ -projected area and (2) CatWalk ™ -max contact area and print area displayed no correlation with punctate allodynia (Fig. A, B). In addition, a significant correlation was detected with CatWalk ™ -mean intensity ( n = 22, P = 0.050) (Fig. G). Thus, correlation statistics indicated that the coordination and intensity data, but not the area data, were correlated with the mechanical hypersensitivity in CFA rats. Fig. 6 Coordination data are pain-related gait parameters in the CFA model. A Correlation analysis between mechanical threshold (x-axis) and DigiGait-swing (left y-axis) and DigiGait-duty cycle (right y-axis). Linear regression, ** P < 0.01. B Correlation analysis between mechanical threshold (x-axis) an...
A previous study in rats with chronic arthritis showed that 9 mg/kg subcutaneous (s.c.) tramadol slightly reduced the mechanical allodynia, 29 mg/kg s.c. tramadol induced moderate alleviation of punctate allodynia and joint hyperalgesia, and 88 mg/kg s.c. completely rescued these sensory abnormalities to baseline level at 30 min post-administration [ ]. In a sodium monoiodoacetate-induced osteoarthritis model, 100 mg/kg oral tramadol (2 h prior to tests) normalized the CatWalk ™ -max contact area and swing speed [ ]. Taken together, these data led us to select the dose of 30 mg/kg tramadol, and the behavioral assessments were performed during 1 h-2 h after i.p. tramadol injection.
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"text": "Furthermore, correlation analyses were conducted between mechanical threshold and each gait parameter in DigiGait ™ and CatWalk ™. For the coordination data, in both systems, strong correlations were found with duration of swing ( n = 22, DigiGait ™: P = 0.003; CatWalk ™: P = 0.003) and duty cycle ( n = 22, DigiGait ™: P = 0.002; CatWalk ™: P = 0.130) (Fig. A, B). For area data, (1) DigiGait ™ -projected area and (2) CatWalk ™ -max contact area and print area displayed no correlation with punctate allodynia (Fig. A, B). In addition, a significant correlation was detected with CatWalk ™ -mean intensity ( n = 22, P = 0.050) (Fig. G). Thus, correlation statistics indicated that the coordination and intensity data, but not the area data, were correlated with the mechanical hypersensitivity in CFA rats. Fig...."
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"identifier": "10.1007/s12264-018-00331-y"
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