Pretest, conditioning sessions, and test all occurred at the same

Pretest, conditioning sessions, and test all occurred at the same time of day (± 1 h) for each hamster. VS and cocaine were used as stimuli. To test for a CPP for

VS, 22 sexually naïve adult and 18 juvenile hamsters were assigned to control and experimental groups, n = 9–11. To reduce the number of cohorts required and prevent exposing control animals to the smell of the stimuli, control selleck animals were housed in a separate but similar vivaria in which the dark phase began at 08:00 h and testing at 09:00 h. In total 10 conditioning sessions occurred, including five no-stimulus and five stimulus-paired sessions. Including the pretest and test, the experiment took place over 12 consecutive days, from P20 to 31 for juvenile animals and P63–69 to 74–80 for adult animals. An hour before use, VS were collected from 30 females and mixed together to total approximately 500 μL. VS are composed of both non-volatile and volatile components, and both have been shown to have behaviorally relevant properties (Petrulis, 2009). Thus, to ensure exposure to both non-volatile

and volatile components of VS immediately prior to and Romidepsin concentration for the duration of the training session, VS were delivered in two ways. Approximately 15 μL of VS was applied to water-moistened cotton gauze packed into a 2-mL Eppendorf tube, one tube for each male. Immediately before testing, the tube was placed out of reach from the male at the top of the back wall in the initially non-preferred compartment in VS-paired conditioning sessions for the VS group. Empty Eppendorf tubes were used for the control group in all conditioning sessions and for the VS group in the no-stimulus conditioning sessions. To ensure exposure to non-volatile components of VS, the remaining approximately 200 μL BIBF-1120 of VS was mixed with 1 mL of mineral oil, and approximately 10 μL of this mixture was applied with

a metal spatula directly onto the nose of hamsters in the VS group immediately before being placed in the VS-paired compartment. Only the VS group was present and all were restrained to their VS-paired compartment when VS was present in the behavior testing room, thus eliminating any concerns about odor diffusion and non-specific conditioning. Clean oil was applied to the nose of hamsters in the control group for all conditioning sessions and in the VS group for no-stimulus conditioning sessions. One hour after completion of the CPP test, hamsters were killed with an overdose of sodium pentobarbital (150 mg/kg, i.p.) and a terminal blood sample was collected via cardiac puncture for radioimmunoassay of circulating plasma testosterone. To test for a CPP for cocaine, 16 juvenile hamsters were assigned to control and experimental groups (n = 8).

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