wer than that in both the IC and SC groups, but no difference between the IC and SC groups was found. Following repeated nicotine administration, the level of pDARPP-32 Thr34 in PFC was increased in the EC = 8.32, p,0.05) and IC = 3.12, p,0.05) compared to the respective saline controls. No difference between SC-Sal and SC-Nic groups was found. In NAc, two-way ANOVA on the AT 7867 manufacturer levels of pDARPP-32 Thr34 revealed a main effect of housing condition = 4.21, p,0.05), and no significant effect of treatment or their interaction. EC rats exhibited decreased basal pDARPP-32 Thr34 level compared to the IC and SC rats. Repeated nicotine significantly increased pDARPP-32 Thr34 level in the EC rats, but not in IC and SC rats. In striatum, no difference in pDARPP-32 Thr34 level was found among EC, IC and SC rats with nicotine or saline injection. Repeated Nicotine Administration Differentially Regulated Phosphorylation of CREB in EC, IC, and SC Rats We examined whether environmental enrichment changed CREB and pCREB in the PFC, NAc, and striatum in the EC, IC, and SC groups. As shown in 7 Enriched Environment Regulates Signaling Proteins total CREB were found in these regions among the groups. With respect to the ratio of pCREB /CREB in the PFC, a main effect of housing condition = 21.22, p, 0.001) and treatment = 
PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22212565 98.64, p, 0.001), and a significant housing condition 6 treatment interaction = 4.69, p, 0.05) were found. Post hoc analysis revealed that the ratio of pCREB /CREB was lower in EC-Sal than in IC-Sal = 11.44, p, 0.05) and SC-Sal = 58.00, p,0.001), indicating that environmental enrichment decreases the basal levels of pCREB. Repeated nicotine administration significantly increased pCREB levels in PFC of EC = 88.57, p,0.001), IC = 29.57, p,0.01), and SC = 21.35, p,0.01) compared to the respective saline control groups. With respect to the ratio of pCREB /CREB in the NAC, a main effect of housing condition = 9.34, p,0.05) was found. There was no significant effect of treatment and their interaction. The ratio of pCREB /CREB was lower in EC-Sal than in IC-Sal = 5.92, p,0.05) and SCSal = 4.89, p,0.05). Repeated nicotine administration increased the level of pCREB in EC-Nic rats = 6.71, p,0.05) but not in IC-Nic and SC-Nic rats. In striatum, no differences in pCREB and total CREB levels were found among the EC, IC and SC rats with nicotine or saline injection. Alterations of Locomotor Behavior were Associated with pDARPP-32 Thr34 Levels in PFC To determine whether the basal level of DARPP-32 activity was associated with the results of behavior tests, the correlation of locomotor activity and DARPP-32 activity was examined. Discussion The current findings demonstrate that an enriched housing environment alters the levels of phosphorylated DARPP-32 and Enriched Environment Regulates Signaling Proteins CREB under control conditions and following nicotine administration. Specifically, the effects of enrichment on activity of DARPP-32 and CREB are robustly found in PFC relative to NAc and striatum. The fact that the basal phosphorylation state of DARPP-32 at Thr34 site in PFC is positively correlated with locomotor activity in EC, IC, and SC rats under saline control conditions, suggests that the PFC DARPP-32 phosphorylation at Thr34 may play an important role in enriched environment-induced changes in locomotion. Acute nicotine injection produces increased levels of pDARPP-32 Thr34 in EC rats in a dose-dependent manner. Although repeated nicotine administ
