r34. Compared to IC rats, EC rats exhibit decreased expression and functioning of D1 receptors in PFC, which may cause lower dopaminergic tone under basal condition in EC rats compared to IC rats. On the other hand, environmental enrichment enhances excitatory glutamatergic synaptic transmission in cortex, which may induce dephosphorylation of Thr34 by activation NMDA receptors. Therefore, an altered balance of D1/cAMP/PKA signaling and glutamate transmission in PFC may cause a difference in the basal levels of pDARPP-32 Thr34 between EC and IC rats. Acute nicotine regulated the phosphorylation levels of DARPP32 at Thr34 and Thr75 sites in a dose-dependent manner. While acute nicotine only increased Thr34 levels in NAc of EC rats and had no effects on Thr75, the high dose of nicotine increased Thr34 levels in PFC and NAc levels in EC rats. However, nicotine obviously increased Thr34 levels in all regions and Thr75 levels in striatum in EC rats, suggesting an enhanced maximal action of nicotine on the phosphorylation of DARPP-32. Under in vitro conditions, nicotine at a low concentration PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22212322 decreases Thr34 levels, whereas nicotine at a high concentration increases Thr34 and decreases Thr75 levels. Although systemic acute nicotine increased phosphorylation of both Thr34 and Thr75 in mouse striatum 15 min after injection, another report showed no effects on Thr34 and Thr75 in striatum and NAc of rats 20 min after a single injection of nicotine . Thus, nicotine-mediated regulation of DARPP-32 activity is largely dependent on the species, dosage, route of administration, and the time needed to harvest brains. Importantly, in the current study, nicotine produced increased pDARPP-32 Thr34 in EC rats, which may be caused by an intrinsic difference in basal levels of Thr34 between EC and IC rats. The action of nicotine on Thr34 is regulated by activation of the D1 receptor-mediated PKA pathway, and this cascade contributes to nicotine-induced motivation. The current results show that the behaviorally-relevant dose of nicotine produce hyperactivity in EC and IC rats, but hypoactivity in SC rats on Day 1. Thus, the differential regulatory effects on pDARPP-32 Thr34 levels in response to acute nicotine in EC and IC rats may play a role in the differential locomotor response to nicotine between EC and IC rats. Repeated nicotine ML 176 price administration eliminated the basal difference in pDARPP-32 Thr34 observed between the EC and IC rats and increased pDARPP-32 Thr34 in the PFC of EC rats relative to IC rats. Nicotine also elicited Thr34 increases in the NAc of EC rats, despite being not as robust as in the PFC. This implicates that the processes mediating the lower basal levels of pDARPP-32 Thr34 in the PFC of EC rats do not prevent repeated nicotine stimulation from regulating DARPP-32 signaling. Rather, compared to their respective saline controls, the magnitude of change in nicotine-induced Thr34 level in PFC is greater in EC than in IC and SC rats. DA D1 receptor activation has been demonstrated to increase Thr34 levels 
and it is possible that the greater increase in nicotine-induced Thr34 levels in PFC of EC rats may represent a compensatory D1 receptor-mediated down-regulation in response to nicotine-stimulated enhancement of DA transmission. While the current results show that no effects of repeated nicotine stimulation on pDARPP-32 Thr34 levels in striatum were found in EC, IC, and SC rats, a previous study has shown that repeated nicotine produces a cl