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In this study, SCR mice showed significant differences in the contextual tests.
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Contextual test (5 min) was performed 24 h after training, in the same chamber.
For the contextual test, mice were returned to the same training chamber for 5 min without an electric shock.
When the conditioned stimulus (context) was presented 24 h after conditioning (context testing), SCR mice showed reduced levels of freezing [Fig. 1B, Contextual test after conditioning, p = 0.0266, f(1,38) = 5.324].
A cued test was conducted in a different environment 2 h after the contextual test using a white plastic box with a brightness of 50 lux, background white noise of 60 dB, and presence of the smell of alcohol.
In the contextual test (Figure 6C, right), performed 24 h after training, PC3/Tis21−/− mice spent a significantly smaller amount of time in freezing behavior, compared to control mice (one-way ANOVA, F 1,22) = 5.02; p = 0.03).
A contextual test was conducted 24 (day 1), 48 (day 2), and 72 h (day 3) after conditioning in the same chamber without the presentation of the tone or footshock.
To evaluate contextual fear learning, test subjects were returned to the freeze monitor (i.e., same context) 24 hr after the training procedure described above, and freezing behavior was scored for 5 min. To evaluate cued fear learning, the animals were placed in a different context (novel odor, lighting, cage floor, and visual cues) 24 hours following contextual testing.
After an initial training period, the mice were either returned to the test chamber for 5 min with no auditory cue (contextual test) or placed in a similar chamber with an auditory cue (memory test) 1 and 24 h after the training session.
The effect of time was significant during contextual testing after 24 h (Fig. 1B) in WT but not in SCR [WT; p = 0.0013, SCR ; p = 0.1943], while the time interaction was not significant between 2 genotypes [p = 0.1527, f(4,152) = 1.701].
It is therefore possible that decreased anxiety-like behavior in CaMKIV KO mice shifted their coping behavior from a freezing response to a fleeing response and then resulted in lower non-specific freezing in the contextual test and during pre-tone and tone periods in the altered context chamber on Day 28.
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