We employed an individually titrated conditioning stimulus, namely the submaximal effort tourniquet test (SETT) [33], and applied three different types of test-stimuli.
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Naïve mice were placed individually into the conditioning box.
On the day of testing (Day 1 to 7), mice were placed individually into the same conditioning chamber for 6 min. The fear conditioned freezing behavior was scored as previously described [13], [26].
Animals were individually placed in standard conditioning boxes (see above).
Each rat was placed individually in the operant conditioning chamber (30×22×24 cm; Med Associates, St . Albans VT) equipped with two retractable levers (45 mm wide ×2 mm thick, protruding 20 mm from the wall) below cue lights on a side wall (Fig. 1B) and a standard tone generator (2900 Hz, Sonalert, Med Associates).
24 hr later, mice were placed individually into the same conditioning chamber for 6 min.
When test mice were placed individually in a fear-conditioning arena and they were presented with the CS-only (the CS was 30-s tone) for 10 consecutive trials, their freezing responses were minimal and strain-independent (Fig. 2a; effect of genotype, F[1], [20] = 1.60, P = 0.22), indicating that presentation of the CS was not salient without conditioning.
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It was speculated that this was perhaps a result of their Japanese cultural conditioning to appear individually modest while reverential to peers.
