Divided into two stages of training and testing Â
(1) Training phase (first day) Â
1. Debug the instrument to ensure that the grille floor is irritated by current, the loudspeaker is audible, and the current intensity and sound intensity (decibel) are recorded separately. Â
2. Rats were placed in a conditional fear box for 2 min (phase A) and the freezing time of the animals for the first 2 min was recorded as a baseline. Â
3. Then add the click sound, 80Db, 30s (B phase). Â
4. Immediately after the electric shock, 0.35mA, 2s (C phase), complete a round of training. If multiple rounds of training are required, repeat A, B, and C three-phase training. Â
5. The animal's stagnation time (s) was recorded throughout the training period to measure the unconditioned fear of the animal. Â
6. Remove the rat from the operating box. Thoroughly clean the box with 70% alcohol or 4% acetic acid for training in the next batch of animals. Â
(b) The test phase is performed on the second day after training, including context testing, altered context or pre-conditioning-stimulus (pre-CS) testing, and conditioning stimulus (conditioning stimulus, CS) Test. There was no electric shock stimulation during the test phase. There are also no auditory conditional stimuli in the association test and change association test, but the operation box used by each animal should be the same as the training phase. Â
1. The association test puts the animals in the box and the computer automatically records the animal's stagnation behavior. If observed by hand, the observer (should not know the animal's treatment or genotype) observes the animal's behavior through the cat's eye (small window) on the door of the box with a static sound stopwatch and scores: stagnation is 1 point, activity (moving) It is 0 points (there is also only the stagnation time recorded). Observe every 10 seconds for a total of 5 minutes, so that 30 possible bouts of possible stagnation (ie, a maximum stagnation score of 30) can be recorded. The animal's contextually conditioned fear was measured using the lag time of the animals recorded in the same operating box recorded at this stage. This is the association test. After the observation, the animals were returned to the cage, and the operation box was cleaned for observation of the next animal. Â
(1) Training phase (first day) Â
1. Debug the instrument to ensure that the grille floor is irritated by current, the loudspeaker is audible, and the current intensity and sound intensity (decibel) are recorded separately. Â
2. Rats were placed in a conditional fear box for 2 min (phase A) and the freezing time of the animals for the first 2 min was recorded as a baseline. Â
3. Then add the click sound, 80Db, 30s (B phase). Â
4. Immediately after the electric shock, 0.35mA, 2s (C phase), complete a round of training. If multiple rounds of training are required, repeat A, B, and C three-phase training. Â
5. The animal's stagnation time (s) was recorded throughout the training period to measure the unconditioned fear of the animal. Â
6. Remove the rat from the operating box. Thoroughly clean the box with 70% alcohol or 4% acetic acid for training in the next batch of animals. Â
(b) The test phase is performed on the second day after training, including context testing, altered context or pre-conditioning-stimulus (pre-CS) testing, and conditioning stimulus (conditioning stimulus, CS) Test. There was no electric shock stimulation during the test phase. There are also no auditory conditional stimuli in the association test and change association test, but the operation box used by each animal should be the same as the training phase. Â
1. The association test puts the animals in the box and the computer automatically records the animal's stagnation behavior. If observed by hand, the observer (should not know the animal's treatment or genotype) observes the animal's behavior through the cat's eye (small window) on the door of the box with a static sound stopwatch and scores: stagnation is 1 point, activity (moving) It is 0 points (there is also only the stagnation time recorded). Observe every 10 seconds for a total of 5 minutes, so that 30 possible bouts of possible stagnation (ie, a maximum stagnation score of 30) can be recorded. The animal's contextually conditioned fear was measured using the lag time of the animals recorded in the same operating box recorded at this stage. This is the association test. After the observation, the animals were returned to the cage, and the operation box was cleaned for observation of the next animal. Â
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