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Table 1.

Treatment regimes

ExperimentTreatments
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–1Determine 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 
ExperimentTreatments
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–1Determine 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 
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