For the novel object recognition test (NORT) and place recognition test (PRT), a PVC box, measuring 80 cm width × 80 cm length with a 50 cm of height, provided by Muromachi Kikai Co., Tokyo, Japan, was used. For NORT and PRT, the rats were allowed to explore the empty box for 10 min for 2 days continuously. On the 3rd day, during the trial phase, the rats were allowed to explore two identical objects (mugs) for 5 min, followed by another 5 min of a retention phase. During the test phase, a new object was replaced with one of the mugs and the explorative time at the novel object by the rats was recorded using the ANY-maze™ Video Tracking System. A similar protocol was used for PRT, however, in PRT, two identical objects were placed in one selected location (north and south) initially. During the test phase, one of the identical objects was transferred to the new location (west). In PRT, the explorative time at the new location by the rats was recorded using the ANY-maze™ Video Tracking System [28 (link)].
Video tracking system
Manufactured by ANY-maze
Sourced in United States
The ANY-Maze Video Tracking System is a software tool designed for automated behavioral analysis. It provides real-time tracking and recording of animal movements within a defined experimental setup.
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Lab products found in correlation
35 protocols using video tracking system
1
Novel and Place Recognition Behavioral Testing in Rats
2
Measuring Social Preference and Novelty in Mice
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The social preference and social novelty assessments were performed as described [26 (link), 27 (link)] with minor modifications. Individual mice were placed in the 3-chambered box and allowed to freely explore the arena during a 10-min habituation period. After the habituation period, an unfamiliar, same-sex mouse of a different genotype (stranger 1) was placed in one of the side chambers under a wire cage. An identical wire cage containing an inanimate object was placed in the opposite chamber. The test mouse was then allowed to explore the entire 3-chambered arena for 10 min. The amount of time spent in each chamber was recorded by the AnyMaze video-tracking system. Following this period, a second unfamiliar, same-sex mouse of a different genotype (stranger 2) was placed into the wire cage previously containing the inanimate object. The test mouse was then allowed to explore the 3-chambered arena for 10 min. The amount of time spent in each chamber was recorded by the AnyMaze video-tracking system. Based on the amount of time spent in each chamber, a ‘sociability index’ and a ‘social novelty index’ was calculated as previously described [27 (link)]. The sociability index was calculated as timestranger/(timestranger + timeobject) × 100. The social novelty index was calculated as timenovel/(timenovel + timefamiliar) × 100.
3
Rotarod Testing for Motor Function
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Rotarod testing was conducted as previously described (41 (link), 42 (link)). The functional test was conducted at P30 and P60 with the ANY-Maze Video Tracking System (ANY-Maze, USA). On the day before the test, the animals were brought to the testing room and allowed to rest for 2 h before testing. After a training session of two consecutive trials before the testing day, the mice were subjected to test sessions with two speed modes: accelerating speeds (range, from 0 to 50 rpm) and fixed speeds (fixed, 40 rpm). Each test mode consisted of two trials on the rotarod, with a maximum of 300 s. Each trial interval lasted more than 1 h. The time each mouse spent on the rotarod was recorded, and the average time of each trial was used for analysis (43 (link)).
4
Open-Field Test for Rat Exploration
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The open-field test was conducted according to a previous methodology [15 (link)]. The rats were acclimated in the testing room and left alone for 30 min before the test. A square box (40 cm [length] × 40 cm [width] × 30 cm [high]) was used to evaluate the exploratory activity of rats. The floor area included both the surrounding and center regions. At the beginning of the trial, each rat was placed in the middle zone. The ANY-Maze Video Tracking System (ANY-Maze, St. Louis, MO, USA) tracked the rat’s movements for 5 min. Moreover, the total traveled distance and the number of lines crossing for each animal were recorded using the software. The grooming time was recorded independently by two experimenters. Both of them were blinded to the group assignments.
5
Morris Water Maze for Mice
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A white circular plastic tank (122 cm in diameter) with walls 76 cm high, filled with water maintained at 22°±2 °C, made opaque by the addition of a nontoxic white paint, was used to perform the Morris water maze test, as previously described.13 (link) Briefly, mice were trained to swim to a submerged Plexiglas platform from 4 different starting points, on a daily basis for a total of 5 days. If they failed to find the platform within 60 s, they were manually guided to the platform and allowed to remain there for 15 s. Mice were trained to reach the training criterion of 20 s (escape latency). Mice were assessed in the probe trial 24 h after the last training session. The probe consisted in a free 180 s swim in the pool without platform. Each animal's performance was monitored using the Any-Maze video tracking system, which provided data for the acquisition parameters (latency to find the platform and distance swam and) and the probe trial parameters (number of entries in the target platform zone of the platform and time in quadrants).
6
Quantifying Repetitive Grooming Behavior
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The test was performed as previously described [27 (link), 29 (link)] to evaluate repetitive grooming tendencies. Mice were individually habituated in an empty test cage for 30 min prior to the test. To amplify natural grooming tendencies, mice were misted with water 3 times at 10 cm distance of the upper-back. Following this misting, the grooming behaviour of each mouse was recorded by the Anymaze video-tracking system for 30 min. The time that each individual mouse spent grooming during this 30-min trial was manually scored by the rater, unaware of the genotype.
7
PTZ-Induced Seizure Evaluation in Mice
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WT and KO littermates implanted for EEG monitoring (see below) were repeatedly injected with unitary doses of PTZ (10 mg/kg intraperitoneally in 0.9% saline) every 10 min and continuously monitored after each injection in a 17 × 17 × 25 cm box equipped with the Anymaze video tracking system. Seizure scoring was conducted as previously reported by Browning and Nelson (1986) (link), and the following parameters were considered: (i) myoclonic jerk, (ii) face and forelimb clonus (iii) whole body clonus with twisting or loss of posture, (iv) running/bouncing clonus, (v) tonus: (tonic flexion and tonic extension) (Browning and Nelson, 1986 (link)). At the end of the observation period, animals were killed humanely by cervical dislocation. Seizure manifestations were recorded by inspection of the videos by two independent observers blind to the genotype. Threshold dose (mg/Kg) for induction of seizures, as well as latency (s) and duration (s) of induced seizures were computed as previously described (McLeod et al., 2013 (link), Rantala et al., 2015 (link)). An integrated measure of seizure propensity was calculated, for each animal, as follows: [seizure duration]/([seizure latency] × [threshold dose]) × 100. The total covered distance and duration of freezing events in the pre-seizure doses were automatically assessed using the Anymaze software.
8
Open Field Anxiety Behavior in Mice
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A second measure of anxiety behaviour included was the open field test. Mice were placed in the corner of a brightly lit (650 lux) box (50 x 50cm x 35cm) and videotaped with AnyMaze Video Tracking System for 5 minutes. Time spent in the pre-defined zones (periphery and center) was recorded. Anxiety and exploratory behaviors were measured and included the amount of time an animal spent in the center or peripheral zone, as well as general measures of motor activity.
9
Elevated Plus Maze for Anxiety Assessment
10
Open Field Test for Anxiety-like Behaviors in Mice
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The open field test was performed to evaluate anxiety-like behaviors in mice as published [20 (link)]. Animals were placed in the center of a PVC arena (40 × 40 × 30 cm) and allowed to explore freely for 5 min. Mouse movements were video-recorded and analyzed using the ANY-maze video tracking system to determine the total travel distances and time spent in the center and four corner zones. The center zone is defined as a square area of 20 × 20 cm in the center of the arena. The outside zone is the area outside of the center zone in the arena.
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