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Before each trial, the subject was given a 5 min acclimation period in the start box.
A 5 min acclimation period commenced the startle session, in which baseline movement was assessed at a background noise level of 70 dB.
All pain tests were performed on each animal in the following order, movement-evoked pain, spontaneous pain (after the 20 min acclimation period), and tactile hypersensitivity.
Patches were visually obscured from one another by plastic barriers during a 30 min acclimation period, during which time fish were also enclosed within a 6 mm mesh cage to prevent predation (as above).
Following a 5 min acclimation period, 4 trial types (5 trials each) were administered in a Latin square design over 7 min: 1) no stimulus; 2) stimulus alone (120 db SPL white noise burst) and prepulses at 3) 70 dB and 4) 76 dB.
Swimming speed was increased every two minutes after a 20 min acclimation period.
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A background noise level of 65 dB was presented for a 10-min acclimation period and continued throughout the test session.
During both the 12-h period in darkness and the 60-min acclimation period before the administration of drugs, at both 6 and 12 weeks, animals of each of the 3 birth groups exhibited similar locomotor activity (data not shown).
After a 5-min acclimation period with 70-dB-background noise (white noise), an 75-, 85-, 85-, 90-, 100-, 110-, or 120-dB white noise stimulus (40 msec duration) was given 8 times to each mouse in the same pseudo-random order at 15 sec intervals.
Fish were then allowed an additional 5-min acclimation period before starting the experiment.
Again, a 7-min acclimation period was followed by a 7-min observation period.
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