Neurobehavior Assessment

The neurobehavioral assessment was carried out following the last day of Cannabis sativa administration (day 22). All neurobehavioral tests were recorded using a digital camcorder and were scored later by two independent trained observers. The following tests were carried out.

The procedure as carried out by Kraeuter et al. and modified by Adelodun et al. was utilized for memory assessment^{18, 19} This was a test designed to evaluate mice’ capacity for learning and recalling spatial information. Three opaque wooden arms formed a Y at a 120° angle from one another, where the tests were conducted. The rats were introduced at the center and allowed to move freely exploring the three arms. The arms of the Y maze were labeled as A, B, and C. The maze was cleaned with alcohol and allowed to dry before subsequent use. The position was maintained throughout the Y maze test of all the rats. Each rat was taken and placed in one of the arms (same arm for all rats) facing the center. After 5 minutes of quiet exploration, each rat was released into the open arms of the Y maze. The numbers of entries into each arm (A, B, and C) and the number of spontaneous alternations were recorded. When an experiment was over, the rats were returned to their original groups.

The percentage of spontaneous alternation was calculated using the following formula:

This was utilized in experimental Wistar rats to assess locomotor and exploratory behaviors. Blanchard et al.’s work was modified for this procedure. ²⁰ Seven rats from each group were chosen and placed in the center of the device, a white open box (72×72×36 cm) with black dividing lines, to assess locomotor and exploratory activity. Ethanol (70%) was used to clean both apparatuses in between tests to prevent olfactory cues. While exploring, the animals were recorded with a camcorder (DNE webcam, Porto Alegre, Brazil) placed at the top and the video was analyzed by independent observers blind to the procedure. The following parameters were scored: lines crossed, center square entries, rearing, and freezing. After each test, each rat was placed back in their respective groups.