Treadmill Locomotion Kinematic Recording
Objective: High-speed videographic analysis of mouse gait and inter-limb coordination across a range of treadmill speeds from 5 to 150 cm/s
Protocol Steps
Animal acclimation to treadmill
Mice were allowed to acclimate quietly on the treadmill lane without movement
Note: Initial familiarization phase before training
View evidence from paper
“First, mice were allowed to acclimate quietly on the lane for 20–30 min”
Initial speed training
Mice were introduced to walk at low speed with electrified grid at minimal intensity to motivate locomotion
Note: Electrified grid set at 0.1 mA minimal intensity; mice learn to walk to avoid grid
View evidence from paper
“They were then introduced to walk at 10–15 cm/s for 5 min. At that stage, the mice kept walking on the treadmill belt to avoid the electrified grid”
Training phase completion
From nine mice trained, six successfully learned to avoid electrified grid; three were excluded from study
Note: Selection criterion: ability to avoid electrified grid during training
View evidence from paper
“Among the group of nine mice used during the training phase, six learned to avoid the electrified grid. The three remaining mice were excluded from the study”
Progressive speed increase training
Mice were walked at increasing speeds until reaching 100 cm/s
Note: Preparation for testing at multiple speeds
View evidence from paper
“Mice were walked at increasing speed. Once they reached 100 cm/s, they were tested 3 times at each speed”
Kinematic recording at multiple speeds
Mice were filmed from left and right sides at 12 different treadmill speeds while walking
Note: Speeds tested: 5, 10, 15, 30, 45, 60, 75, 90, 105, 120, 135, and 150 cm/s; at least 10 contiguous strides recorded per speed
View evidence from paper
“All mice were filmed on the left and right sides by high-frequency (200 frames/s) cameras (Genie HM640, Dalsa Teledyne) during treadmill locomotion. To study inter-limb coordination over a wide range of locomotor speeds, mice were tested at treadmill belt speeds of 5, 10, 15, 30, 45, 60, 75, 90, 105, 120, 135, and 150 cm/s”
Marker placement for trajectory analysis
Reflective markers were glued to specific anatomical landmarks on hindlimbs and forelimbs under anesthesia
Note: Performed under isoflurane anesthesia (2-3%); limbs were shaved before marker placement
View evidence from paper
“Under isoflurane anesthesia (2–3%), limbs were shaved and reflective markers (2 mm diameter) were glued on the iliac crest, hip, knee, ankle, and metatarsophalangeal (MTP) joints and toe for the hindlimb, and on the scapula, shoulder, elbow, and metacarpophalangeal joints and toe for the forelimb”
Kinematic recording at selected speeds with markers
Mice walked at three specific speeds with reflective markers to investigate limb trajectory
Note: Speeds: low (15 cm/s), intermediate (45 cm/s), and high (90 cm/s)
View evidence from paper
“mice walked at low (15 cm/s), intermediate (45 cm/s), and high speeds (90 cm/s) with reflective markers”
Video digitization and analysis
Recorded films were digitized using StreamPix software and analyzed offline
Note: Offline analysis allows detailed kinematic measurements
View evidence from paper
“Films were digitized with StreamPix 6.0 (Norpix) and analyzed offline”