This effect was observed after long-term, but not short-term, withdrawal; however, early acquisition of sensitized reactions was not investigated (Pierce and Kalivas, 1995). requirements of D1R for dopamine-dependent behaviors. Intro Differential gene manifestation within discrete mind areas expands neural coding capacity and diversifies circuit function. This is exemplified in the striatum, where two parallel circuits, the direct and indirect pathway, oppositely regulate thalamocortical loops. These pathways possess a related neuronal cell type, the medium spiny neuron, yet differ dramatically in connectivity, neuropeptide manifestation, and genetic profiles. The balance of circuit activation between the direct and indirect pathway is necessary for several behaviors, including reward processing (Lobo et al., 2010, Beutler et al., 2011). The dopamine D1 receptor (D1R), encoded from the gene, is definitely highly enriched in the direct pathway (Fig. 1= 7) and D1R-NAcShell (= 9) mice. Level bars: by insertion of Cre recombinase were explained previously (Heusner et al., 2008). food and water except during food restriction to 85% of their bodyweight. Generation of AAV-FLEX-D1RGFP, viral injections, and experimental organizations. The adeno-associated computer virus (AAV)-FLEX-D1RGFP was generated by PCR amplification of D1R from genomic DNA (C57BL/6J) using the primers 5-GATATCACCGGTATGGCTCCTAACACTTCTAC-3 and 5-GATATCGCGGCCGCGGTTGAATGCTGTCCGCTGT-3. The 1.3 kb PCR product was subcloned into AM/CBA-FLEX-EGFP-WPRE-bGH in-frame with EGFP. AAV was generated as explained previously (Zweifel et al., 2008). For stereotaxic viral injections, 0.5 l of AAV-FLEX-D1RGFP (titer 1 1012/ml) or control AAV-FLEX-GFP (titer 1 1012/ml) was bilaterally injected into the NAcCore (= 1.0, = +1.3*F, = ?4.25) or NAcShell (= 0.4, = +1.3*F, = ?5.0), = [lambda ? bregma]/4.21. To control for effects of site-specific injections and viral-mediated D1R manifestation in restricted NAc subregions, we generated the following experimental organizations: NAcCore, Het GFP-NAcCore (recombinase manifestation cassette into the open reading frame of the locus (Heusner et al., 2008). This results in selective manifestation of in D1R-containing cells. Mice homozygous for the insertion are null mutants, insertion are indistinguishable from additional D1R knock-out mouse lines (Drago et al., 1994, Xu et al., 1994). To re-express D1R in an PEPA anatomically restricted manner, we generated an AAV vector comprising a Cre-conditional D1R-GFP manifestation cassette (AAV-FLEX-D1RGFP, Fig. 1= 7) displayed little locomotor response to the drug (Fig. 2= 7) showed a strong agonist effect that was indistinguishable from that of heterozygous control organizations (Het GFP-NAcCore, = 7; Het D1R-NAcCore, = 8; two-way repeated-measures ANOVA, genotype time, = 0.0282; Fig. 2= 9) mice also responded to SKF-81297 with significantly improved locomotor activity compared with GFP-NAcShell mutants (= 7; two-way repeated-measures ANOVA, genotype time, 0.0001; Fig. 2= 12; Het D1R-NAcShell, = 13; Fig. 2= 7; Het-D1R, = 8; Mut-GFP, = 7; Mut-D1R, = 7; NAcShell: Het-GFP, = 12; Het-D1R, = 13; Mut-GFP, = 7; Mut-D1R, = 9). = 8; Het-GFP, = 5; Het-D1R, = 5; Mut-GFP, = 6; Mut-D1R, = 6; NAcShell: saline settings, all genotypes, = 9; Het-GFP, = 10; Het-D1R, = 10; Mut-GFP, = 6; Mut-D1R, = 7). 0.05, ** 0.01 for D1R-NAcCore or D1R-NAcShell mice versus D1R mutants, respectively. 0.01, *** 0.001 for D1R mutants versus all other groups. Scale bars, 100 m. Data are demonstrated as means SEM. To further confirm that signaling events downstream of D1R activation are present in D1R-NAcCore and D1R-NAcShell mice, we quantified c-Fos manifestation around the area of viral repair after SKF-81297 administration (7.5 mg/kg; Fig. 2= 6) and control mice (Het GFP-NAcCore, = 5; Het D1R-NAcCore, = 5) showed strong c-Fos induction (one-way ANOVA, 0.0001; Fig. 2= 8) and SKF-81297-treated GFP-NAcCore mutants (= 6) showed negligible c-Fos manifestation (Fig. 2= 7) and control mice (Het GFP-NAcShell, = 10; Het D1R-NAcShell, = 10) also displayed strong induction of c-Fos compared with saline-injected settings (all genotypes, = 9) and SKF-81297-treated GFP-NAcShell mutants (= 6; one-way ANOVA, 0.0001; Fig. 2= 8; Het D1R-NAcCore, = 8), we did not find the head entry rate in D1R-NAcCore mice (= 7) to be significantly above their respective mutant control group (GFP-NAcCore, = 7) during CS demonstration (two-way repeated-measures ANOVA, genotype time, = 0.0222; Fig. 3= 9) also failed to increase their head entry rate significantly above respective mutant settings (GFP-NAcShell, = 7; Fig. 3 0.0001; Fig. 3 0.0001; Fig. 3= 8; Het-D1R, = 8; Mut-GFP, = 7; Mut-D1R, = 7; NAcShell: Het-GFP, = 13; Het-D1R, = 13; Mut-GFP, = 7; Mut-D1R, = 9). 0.01, *** 0.001. Data are demonstrated as means SEM. Sufficiency of D1R in the NAc for instrumental conditioning Conditioned approach to the lever by D1R-NAcCore mice during pavlovian conditioning suggests that these animals have assigned some value to the cue, so we investigated whether they would perform an instrumental response (lever press) to acquire reward. Immediately after pavlovian conditioning, mice were given a simple, fixed.These results PEPA highlight dissociated circuit requirements of D1R for dopamine-dependent actions. Introduction Differential gene expression within discrete brain regions expands neural coding capacity and diversifies circuit function. areas expands neural coding capacity and diversifies circuit function. This is exemplified in the striatum, where two parallel circuits, the direct and indirect pathway, oppositely regulate thalamocortical loops. These pathways possess a related neuronal cell type, the medium spiny neuron, yet differ dramatically in connectivity, neuropeptide manifestation, and genetic profiles. The balance of circuit activation between the direct and indirect pathway is necessary for several behaviors, including incentive processing (Lobo et al., 2010, Beutler et al., 2011). The dopamine D1 receptor (D1R), encoded from the gene, is definitely highly enriched in the direct pathway (Fig. 1= 7) and D1R-NAcShell (= 9) mice. Level bars: by insertion of Cre recombinase were explained previously (Heusner et al., 2008). food and water except during food restriction to 85% of their bodyweight. Generation of AAV-FLEX-D1RGFP, viral injections, and experimental organizations. The adeno-associated computer virus (AAV)-FLEX-D1RGFP was generated by PCR amplification of D1R from genomic DNA (C57BL/6J) using the primers 5-GATATCACCGGTATGGCTCCTAACACTTCTAC-3 and 5-GATATCGCGGCCGCGGTTGAATGCTGTCCGCTGT-3. The 1.3 kb PCR product was subcloned into AM/CBA-FLEX-EGFP-WPRE-bGH in-frame with EGFP. AAV was generated as explained previously (Zweifel et al., 2008). For stereotaxic viral injections, 0.5 l of AAV-FLEX-D1RGFP (titer 1 1012/ml) or control AAV-FLEX-GFP (titer 1 1012/ml) was bilaterally injected into the NAcCore (= 1.0, = +1.3*F, = ?4.25) or NAcShell (= 0.4, = +1.3*F, = ?5.0), = [lambda ? bregma]/4.21. To control for effects of site-specific injections and viral-mediated D1R manifestation in restricted NAc subregions, we generated the following experimental organizations: NAcCore, Het GFP-NAcCore (recombinase manifestation cassette into the open reading frame of the locus (Heusner et al., 2008). This results in selective manifestation of in D1R-containing cells. Mice homozygous for the insertion are null mutants, insertion are indistinguishable from additional D1R knock-out mouse lines (Drago et al., 1994, Xu et al., 1994). To re-express D1R in an anatomically restricted manner, we generated an AAV vector comprising a Cre-conditional D1R-GFP manifestation cassette (AAV-FLEX-D1RGFP, Fig. 1= 7) displayed little locomotor response to the drug (Fig. 2= 7) showed a strong agonist effect that was indistinguishable from that of heterozygous control organizations (Het GFP-NAcCore, = 7; Het D1R-NAcCore, = 8; two-way repeated-measures ANOVA, genotype time, = 0.0282; Fig. 2= 9) mice also responded to SKF-81297 with significantly improved locomotor activity compared with GFP-NAcShell mutants (= 7; two-way repeated-measures ANOVA, genotype time, 0.0001; Fig. 2= 12; Het D1R-NAcShell, = 13; Fig. 2= 7; Het-D1R, = 8; Mut-GFP, = 7; Mut-D1R, = 7; NAcShell: Het-GFP, = 12; Het-D1R, = 13; Mut-GFP, = 7; Mut-D1R, = 9). = 8; Het-GFP, = 5; Het-D1R, = 5; Mut-GFP, = 6; Mut-D1R, = 6; NAcShell: saline settings, all genotypes, = 9; Het-GFP, = 10; Het-D1R, = 10; Mut-GFP, = 6; Mut-D1R, = 7). 0.05, ** 0.01 for D1R-NAcCore or D1R-NAcShell PEPA mice versus D1R mutants, respectively. 0.01, *** 0.001 for D1R mutants versus all other groups. Scale bars, 100 m. Data are demonstrated as means SEM. To further confirm that signaling events downstream of D1R activation are present in D1R-NAcCore and D1R-NAcShell mice, we quantified c-Fos manifestation around the area of viral repair after SKF-81297 administration (7.5 mg/kg; Fig. 2= 6) and control mice (Het GFP-NAcCore, = 5; Het D1R-NAcCore, = 5) showed strong c-Fos induction (one-way ANOVA, 0.0001; Fig. 2= 8) and SKF-81297-treated GFP-NAcCore mutants (= 6) showed negligible c-Fos manifestation (Fig. 2= 7) and control mice (Het GFP-NAcShell, PROM1 = 10; Het D1R-NAcShell, = 10) also displayed strong induction of c-Fos compared with saline-injected settings (all genotypes, = 9) and SKF-81297-treated GFP-NAcShell mutants (= 6; one-way ANOVA, 0.0001; Fig. 2= 8; Het D1R-NAcCore, = 8), we did not find the head entry rate in D1R-NAcCore mice (= 7) to be significantly above their respective mutant control group (GFP-NAcCore, = 7) during CS demonstration (two-way repeated-measures ANOVA,.
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