Delay contextual and cued fear conditioning was conducted using an automated fear-conditioning chamber (Med Associates, St Albans, VT, USA). The conditioning chamber was interfaced to a PC installed with VideoFreeze software (version 1.12.0.0, Med Associates) and enclosed in a sound-attenuating cubicle. Training consisted of a 2-min acclimation period followed by three tone-shock (CS–US) pairings (80 dB tone, duration 30-s; 0.5 mA footshock, duration 1 s; intershock interval 90-s) and a 2.5-min period, during which no stimuli were presented. The environment was well lit (~ 100 lx), with a stainless steel grid floor and swabbed with almond odor cue (prepared from almond extract; McCormick; 1:100 dilution). A 5-min test of contextual fear conditioning was performed 24-hr after training, in the absence of the tone and footshock, but in the presence of 100 lx overhead lighting, almond odor and chamber cues identical to those used on the training day. Cued fear conditioning, conducted 48-hr after training, was assessed in a novel environment with distinct visual, tactile and orange olfactory cues. Overhead lighting was turned off. The cued test consisted of a 3-min acclimation period followed by a 3-min presentation of the tone CS and a 90-s exploration period. Cumulative time spent freezing in each condition was quantified by VideoFreeze software (Med Associates).
Automated fear conditioning chamber
Manufactured by Med Associates
Sourced in United States
The Automated fear-conditioning chamber is a laboratory equipment designed to study fear-related behaviors in experimental subjects. It provides a controlled environment for administering conditioned stimuli and recording the subject's responses.
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5 protocols using automated fear conditioning chamber
1
Delayed Contextual and Cued Fear Conditioning
2
Delay Fear Conditioning Protocol
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Delay contextual and cued fear conditioning was conducted using an automated fear-conditioning chamber (Med Associates, St Albans, VT, USA) as previously described [45 (link), 50 (link), 52 (link)]. Training consisted of a 2-min acclimation period followed by three tone–shock (CS–US) pairings (80-dB tone, duration 30 s; 0.5-mA footshock, duration 1 s; intershock interval, 90 s) and a 2.5-min period during which no stimuli were presented. The environment was well lit (~ 100 lx), with a stainless steel grid floor and swabbed with vanilla odor cue (prepared from vanilla extract; McCormick; 1:100 dilution). A 5-min test of contextual fear conditioning was performed 24 h after training, in the absence of the tone and footshock but in the presence of 100 lx overhead lighting, vanilla odor, and chamber cues identical to those used on the training day. Cued fear conditioning, conducted 48 h after training, was assessed in a novel environment with distinct visual, tactile, and olfactory cues. The cued test consisted of a 3-min acclimation period followed by a 3-min presentation of the tone CS and a 90-s exploration period. Cumulative time spent freezing in each condition was quantified by VideoFreeze software (Med Associates).
3
Cued and Contextual Fear Conditioning in Rats
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Cued and contextual fear conditioning assays were initiated at the tunnel facility in 9.5- and 14.5-month-old animals using an automated fear conditioning chamber (MedAssociates, Inc.), as previously described (Berg et al. 2020 (link)). Briefly, rats were trained on day one with three tone-shock pairings of 30 s of white noise at 80 dB and a 2-s foot shock of . Contextual conditioning was assessed on day 2, and cued conditioning on day 3. Average motion index was measured automatically using VideoFreeze® software (version 2.7; MedAssociates).
4
Automated Fear Conditioning for Learning and Memory
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At PND 43 ± 1, learning and memory were assessed using an automated fear conditioning chamber (Med Associates) following methods previously described (Copping et al., 2017 (link); Adhikari et al., 2019 (link); Berg et al., 2020b (link)). On day 1, rats were trained via exposure to a series of three noise-shock (conditioned stimulus-unconditioned stimulus; CS-US; 80 dB white noise, 0.7 mA foot shock) pairings inside a sound-attenuated chamber with specific visual, tactile, and odor cues. On day 2, contextual memory was tested by placing each subject back inside the training environment (no noise or foot shock occurred). On day 3, cued memory was evaluated by placing subjects into a novel context with altered visual, tactile, and odor cues. Following a period of exploration, the white noise CS was presented for 3 min. Time spent freezing was measured using VideoFreeze software (Med Associates).
5
Contextual and Cued Fear Conditioning
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Mice were examined in conditioned fear across three days as previously described36 (link)37 (link). On day 1, mice were placed into an automated fear-conditioning chamber (Med Associates) and after 2 min a 72 dB, 2900 HZ tone (CS) was presented for 30 sec that terminated with a 2-sec 0.4 mA scrambled foot-shock (US). Mice remained in the chamber for additional 30 sec before they were returned to their home cages. Twenty-four hr following conditioning, the mice were examined for contextual fear. The animals were returned to the same chamber in which they were conditioned, in the absence of the CS and US for 5-min. On the third day, mice were tested for cued fear conditioning. Mice were placed into a novel chamber and after 2 min, then the CS was presented for 3 min. No US was given. Time spent in freezing was scored by trained observers blinded to the genotype of the animal using Observer software (Noldus Information Technologies) and expressed as the percent time freezing relative to the duration of the interval (pre-CS or CS for cued testing) or the test (context). Freezing was defined as the absence of movement, aside from that required for respiration.
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