Behavioral tests were performed 1 week and 2 weeks after injury, to assess changes in motor functions and coordination, compared to evaluations performed 1 week prior to CCI TBI. All tests were performed during the animals’ light phase; cages were transported to testing rooms at least 30 min prior to testing.
Beam Walking Test (BWT): A BWT was used to assess CCI-induced deficits in fine motor coordination. Mice have a preference for a darkened enclosed environment, as compared to an open illuminated one. Each animal was placed in darkened goal box for a 2 min habituation and then the trial began from the other (light) end of the beam. The beam was constructed with the following dimensions: 1.2 cm (width) × 91 cm (length). The time taken for each animal to traverse the beam to reach the dark goal box and the immobility time spent between the moment when they were initially placed on the beam and when they started walking were documented. Five trials were recorded for each animal before CCI and at 1 and 2 weeks after CCI. The mean times to traverse the beam and the immobility times were calculated, and a plot was generated to evaluate treatment effects; these times were then used for statistical analysis.
Gait analysis: For the gait analysis, mice were tested on a fixed-speed treadmill apparatus (DigiGait; Mouse Specifics). Mice were habituated to the apparatus for 1 min, and then given a 1-min run at 5 cm/s. Following a 1-min rest, the treadmill speed was increased to 15 cm/s. Video was collected at high speed from a ventrally placed camera, and 3–5 s of representative gait video was selected by an experienced but blinded user for automated analysis.
Beam Walking Test (BWT): A BWT was used to assess CCI-induced deficits in fine motor coordination. Mice have a preference for a darkened enclosed environment, as compared to an open illuminated one. Each animal was placed in darkened goal box for a 2 min habituation and then the trial began from the other (light) end of the beam. The beam was constructed with the following dimensions: 1.2 cm (width) × 91 cm (length). The time taken for each animal to traverse the beam to reach the dark goal box and the immobility time spent between the moment when they were initially placed on the beam and when they started walking were documented. Five trials were recorded for each animal before CCI and at 1 and 2 weeks after CCI. The mean times to traverse the beam and the immobility times were calculated, and a plot was generated to evaluate treatment effects; these times were then used for statistical analysis.
Gait analysis: For the gait analysis, mice were tested on a fixed-speed treadmill apparatus (DigiGait; Mouse Specifics). Mice were habituated to the apparatus for 1 min, and then given a 1-min run at 5 cm/s. Following a 1-min rest, the treadmill speed was increased to 15 cm/s. Video was collected at high speed from a ventrally placed camera, and 3–5 s of representative gait video was selected by an experienced but blinded user for automated analysis.
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