After dividing the rats into a control, or con, and model, or mod, group, house them in individual cages. For the tail-clamp method, secure the tail of a rat and measure the clamping time for one minute. In the water deprivation method, remove the feeding bottle from the cage and record the rat for 24 hours.
For the food deprivation method, remove the food from the cage and record the rat for 24 hours. For cold stimulation at four degrees Celsius, place a rat in a bucket of cold water and monitor its activity for five minutes. Then dry the rat with a blower and return it to its original cage.
To administer 45 degrees Celsius heat stress, place the rat in an oven and monitor for five minutes. For the inversion of the light-dark cycle, wrap the cage in a black cloth for one hour to stimulate daytime darkness. Subsequently illuminate the cage for 12 hours at night to mimic daylight and record the data for 24 hours.
For the wet padding experiment, pour 200 milliliters of water into a cage containing 100 grams of padding. Place the rat in the wet cage and monitor for 24 hours. After drying the rat as demonstrated earlier, return it to a cage containing fresh wood shavings.
In the cage incline method, place the rat in a cage angled at 45 degrees against a wall and monitor it for 24 hours. For high speed shaking, place the rat in the mechanical shakers set at 160 RPM, and record data for 15 minutes. After applying stressors, transfer the mod group cage from the CUMS room to the housing room.
