Treatment regimes
| Experiment . | . | . | Treatments . | . | . | . |
|---|---|---|---|---|---|---|
| Experiment 1 | Day 1 | Day 2 | ||||
| Conditioned place preference in response to a single exposure | (i) Determine baseline preference over a 2 min period | Determine conditioned place preference over a 2 min period | ||||
| (ii) 20 min conditioning ± nicotine (0–300 μmol l–1) or ethanol (0–264 mmol l–1) | ||||||
| Experiment 2 | Day 1 | Days 2 and 3 | Day 4 | |||
| Conditioned place preference following three consecutive conditioning sessions | (i) Determine baseline preference over a 2 min period | (i) 20 min conditioning ± nicotine (0–300 μmol l–1) | Determine conditioned preference over a 2 min period | |||
| (ii) 20 min conditioning ± nicotine (0–300 μmol l–1) | ||||||
| Experiment 3 | Day 1 | Days 2 and 3 | Days 4–31 | Day 32 | ||
| Conditioned place preference following 4 weeks of daily conditioning with either 30 μmol l–1 nicotine or 175 mmol l–1 ethanol. N=35 for each treatment group | (i) Determine baseline preference over a 2 min period | (i) 20 min conditioning in absence of any drug | (i) 20 min conditioning ± either 30 μmol l–1nicotine or 175 mmol l–1 ethanol | (i) Determine conditioned place preference over a 2 min period | ||
| (ii) 20 min conditioning to apparatus in absence of any drug | ||||||
| Experiment 4 | Day 1–32 as for experiment 3 above. N=18–20 for each treatment group | Day 33 | ||||
| Conditioned place preference despite an adverse stimulus | (i) Determine place preference despite an adverse stimulus | |||||
| Experiment 5 | Day 1–31 as for experiment 3 above. N=35 for each treatment group | Day 32 | Days 32–37 | Day 38 | Days 39–52 | Day 53 |
| Conditioned place preference following a period of abstinence | (i) Determine place preference for each fish over a 2 min period | No treatment. Fish maintained in aquarium | (i) Determine place preference for each fish over a 2 min period | No treatment. Fish maintained in aquarium | (i) Determine place preference for each fish over a 2 min period | |
| (ii) 10–12 fish selected randomly from each group and assessed for place preference despite an adverse stimulus prior to sacrifice | (ii) 10–12 fish selected randomly from each group and assessed for place preference despite an adverse stimulus prior to sacrifice | (ii) Remaining 10–12 fish from each group assessed for place preference despite an adverse stimulus prior to sacrifice. Brains dissected and stored at minus 70°C prior to preparation of RNA for microarray analysis |
| Experiment . | . | . | Treatments . | . | . | . |
|---|---|---|---|---|---|---|
| Experiment 1 | Day 1 | Day 2 | ||||
| Conditioned place preference in response to a single exposure | (i) Determine baseline preference over a 2 min period | Determine conditioned place preference over a 2 min period | ||||
| (ii) 20 min conditioning ± nicotine (0–300 μmol l–1) or ethanol (0–264 mmol l–1) | ||||||
| Experiment 2 | Day 1 | Days 2 and 3 | Day 4 | |||
| Conditioned place preference following three consecutive conditioning sessions | (i) Determine baseline preference over a 2 min period | (i) 20 min conditioning ± nicotine (0–300 μmol l–1) | Determine conditioned preference over a 2 min period | |||
| (ii) 20 min conditioning ± nicotine (0–300 μmol l–1) | ||||||
| Experiment 3 | Day 1 | Days 2 and 3 | Days 4–31 | Day 32 | ||
| Conditioned place preference following 4 weeks of daily conditioning with either 30 μmol l–1 nicotine or 175 mmol l–1 ethanol. N=35 for each treatment group | (i) Determine baseline preference over a 2 min period | (i) 20 min conditioning in absence of any drug | (i) 20 min conditioning ± either 30 μmol l–1nicotine or 175 mmol l–1 ethanol | (i) Determine conditioned place preference over a 2 min period | ||
| (ii) 20 min conditioning to apparatus in absence of any drug | ||||||
| Experiment 4 | Day 1–32 as for experiment 3 above. N=18–20 for each treatment group | Day 33 | ||||
| Conditioned place preference despite an adverse stimulus | (i) Determine place preference despite an adverse stimulus | |||||
| Experiment 5 | Day 1–31 as for experiment 3 above. N=35 for each treatment group | Day 32 | Days 32–37 | Day 38 | Days 39–52 | Day 53 |
| Conditioned place preference following a period of abstinence | (i) Determine place preference for each fish over a 2 min period | No treatment. Fish maintained in aquarium | (i) Determine place preference for each fish over a 2 min period | No treatment. Fish maintained in aquarium | (i) Determine place preference for each fish over a 2 min period | |
| (ii) 10–12 fish selected randomly from each group and assessed for place preference despite an adverse stimulus prior to sacrifice | (ii) 10–12 fish selected randomly from each group and assessed for place preference despite an adverse stimulus prior to sacrifice | (ii) Remaining 10–12 fish from each group assessed for place preference despite an adverse stimulus prior to sacrifice. Brains dissected and stored at minus 70°C prior to preparation of RNA for microarray analysis |