Journal: bioRxiv

Article Title: Arrestin-3 promotes locomotor sensitization to psychostimulants via JNK signaling in nucleus accumbens

doi: 10.64898/2026.04.28.719936

Figure Lengend Snippet: ( A , B ) WT mice were injected bilaterally into the CPu as with LV encoding VE or VE-Arr3. ( A ) Total AMPH-induced locomotion measured for 90 min after administration of the drug. ( B ) The basal and AMPH-induced locomotor activity on Day 3 for the entire testing period of 30 min (basal activity) and 90 min (AMPH-induced activity) in the groups shown in A . The data were analyzed by two-way repeated measure ANOVA with Group (Genotype + injected virus) as between group and Day or Bin as within group factor. Significance values next to brackets refer to the F value differences between Groups across Days or Bins. * - p<0.05 according to two-tailed Student’s t-test for the Challenge Day (Day 15). ( C , D ) WT mice hemizygous for Cre under D1R promotor (D1R-Cre) were injected into the CPu with adenoviruses encoding double-floxed open reading frame-inverted Venus (dfVE) or WT Arr3 (dfArr3). ( C ) Total AMPH-induced locomotion measured for 90 min after administration of the drug. Statistical analysis was the same as in A . ( D ) The basal and AMPH-induced locomotor activity on Day 4 for the entire testing period of 30 min (basal activity) and 90 min (AMPH-induced activity) in the groups shown in C . Statistical analysis was the same as in B . ( E , F ) WT mice hemizygous for Cre under A2A adenosine receptor promotor (A2A-Cre) were injected into the CPu dfVE or dfA3WT). ( E ) Total AMPH-induced locomotion measured for 90 min after administration of the drug. Statistical analysis was the same as in A . ( F ) The basal and AMPH-induced locomotor activity on Day 5 for the entire testing period of 30 min (basal activity) and 90 min (AMPH-induced activity) in the groups shown in E . ( G ) The expression of Arr3 in the CPu of WT mice following injection of LV-HA-Arr3 of Cre-dependent AAV-VE-Arr3 detected with anti-Arr3 antibody. Representative Western blot includes the standard samples to indicate the position ofthe bands: purified Arr3, ng/lane; VE-Arr3 or HA-Arr3 expressed in HEK DKO cells. ( H ) The expression of exogenous VE-Arr3 driven by Cre-dependent AAV in D1R-Cre and A2AR-Cre mice detected with anti-GFP antibody. Upper panel: representative Western blot; lower panel – quantification of the Western blot data (N=7).

Article Snippet: Mice expressing Cre recombinase under control of D1R promoter (D1RCre) were purchased from the Jackson Laboratory (B6.FVB(129S6)-Tg(Drd1a-cre)AGsc/KndlJ strain on C57Bl/6J-congenic background).

Techniques: Injection, Activity Assay, Virus, Two Tailed Test, Expressing, Western Blot, Purification

Journal: bioRxiv

Article Title: Arrestin-3 promotes locomotor sensitization to psychostimulants via JNK signaling in nucleus accumbens

doi: 10.64898/2026.04.28.719936

Figure Lengend Snippet: ( A , B ) A3KO mice hemizygous for Cre under D1R promotor (D1R-Cre) were injected into the nucleus accumbens (NAc) with AAVs encoding double-floxed open reading frame-inverted Venus (dfVE) or WT Arr3 (dfArr3). ( A ) Total AMPH-induced locomotion measured for 90 min after administration of the drug. ( B ) The basal and AMPH-induced locomotor activity on Day 5 for the entire testing period of 30 min (basal activity) and 90 min (AMPH-induced activity) in the mice shown in A . The data were analyzed by two-way repeated measure ANOVA with Group (Genotype + injected virus) as between group and Day or Bin as within group factor. Significance values next to brackets refer to the differences between Groups across Days or Bins. The p value in A applies to both WT-VE -A3KO-VE and A3KO-Arr3 - A3KO-VE. * - p<0.05 to WT-VE, a - p<0.05 to A3KO-Arr3 according to Bonferroni’s post-hoc test for the Challenge Day (Day 15). ( C , D ) A3KO mice hemizygous for Cre under A2A adenosine receptor promotor (A2A-Cre) were injected into NAc with dfVE or dfArr3. ( C ) Total AMPH-induced locomotion measured for 90 min after administration of the drug. Statistical analysis was the same as in A . ( D ) The basal and AMPH-induced locomotor activity on Day 5 for the entire testing period of 30 min (basal activity) and 90 min (AMPH-induced activity) in the mice shown in C . Statistical analysis was the same as in A, B . In C and D , the p values apply to the differences A3KO-VE – WT-VE and A3KO-Arr3 - WT-VE. * - p<0.05 to WT-VE. ( E ) –The expression of exogenous VE-Arr3 in NAc driven by D1R-Cre and A2AR-Cre detected by immunohistochemistry. The sections were co-stained for the marker of MSNs FOXP1 (red). ( F ) The level of Cre driven expression of VE-Arr3 in comparison with the endogenous Arr3 measured by Western blot with anti-Arr3 antibody. Upper panel: representative western blot; lower panel – quantification of the Western blot data (N=8-10).

Article Snippet: Mice expressing Cre recombinase under control of D1R promoter (D1RCre) were purchased from the Jackson Laboratory (B6.FVB(129S6)-Tg(Drd1a-cre)AGsc/KndlJ strain on C57Bl/6J-congenic background).

Techniques: Injection, Activity Assay, Virus, Expressing, Immunohistochemistry, Staining, Marker, Comparison, Western Blot

Journal: bioRxiv

Article Title: Arrestin-3 promotes locomotor sensitization to psychostimulants via JNK signaling in nucleus accumbens

doi: 10.64898/2026.04.28.719936

Figure Lengend Snippet: ( A , B ) A3KO mice hemizygous for Cre under D1R promotor (D1R-Cre) were injected into NAc with AAVs encoding double-floxed open reading frame-inverted Venus (dfVE) or WT Arr3 (dfArr3). ( A ) Total COC-induced locomotion measured for 90 min after administration of the drug. ( B ) The basal and COC-induced locomotor activity on Day 5 for the entire testing period of 30 min (basal activity) and 90 min (AMPH-induced activity) in mice shown in A . The data were analyzed by two-way repeated measure ANOVA with Group (Genotype + injected virus) as between group and Day or Bin as within group factor. Significance values next to brackets refer to the differences between Groups across Days or Bins. * - p<0.05 to WT-VE, a - p<0.05 to A3KO-Arr3 according to Bonferroni’s post-hoc test for the Challenge Day (Day 15). ( C , D ) A3KO mice hemizygous for Cre under A2A adenosine receptor promotor (A2A-Cre) were injected into NAc with dfVE or dfArr3 AAVs. ( C ) Total COC-induced locomotion measured for 90 min after administration of the drug. Statistical analysis was the same as in A . ( D ) The basal and COC-induced locomotor activity on Day 5 for the entire testing period of 30 min (basal activity) and 90 min (COC-induced activity) in the mice shown in C . Statistical analysis was the same as in A, B . * - p<0.05 to WT-VE.

Article Snippet: Mice expressing Cre recombinase under control of D1R promoter (D1RCre) were purchased from the Jackson Laboratory (B6.FVB(129S6)-Tg(Drd1a-cre)AGsc/KndlJ strain on C57Bl/6J-congenic background).

Techniques: Injection, Activity Assay, Virus

Journal: bioRxiv

Article Title: Arrestin-3 promotes locomotor sensitization to psychostimulants via JNK signaling in nucleus accumbens

doi: 10.64898/2026.04.28.719936

Figure Lengend Snippet: (A , B) A3KO D1R-Cre mice were injected with Cre-dependent AAV encoding Venus or VE-T16-F10V and tested for drug-induced locomotion for 7 consecutive days and then for 1 day after a 7-day break. (A) AMPH-induced locomotion. (B) COC-induced locomotion. Bracket shows significant difference between the A3KO-VE and A3KO-VE-T16-F10V groups. ( C , D) Immunohistochemical detection of Venus or VE-T16-F10V in the NAc. ( C ) The sections were stained with anti-GFP antibody and co-stained for the marker of MSNs striatal neurons FOXP1. (D) Adjacent sections through NAc stained for GFP to detect Venus or VE-T16F10V co-stained with anti-Cre antibody. (E , F , G) JNK3 was immunoprecipitated with anti-JNK3 antibody and blotted for doubly phosphorylated (active) JNK3 (ppJNK3). (E) Representative western blots showing immunoprecipitation. (F) Quantification of active JNK3 in the NAc following the AMPH treatment and testing (N=6-7). (G) Quantification of active ppJNK3 in NAc following the COC testing (N=7).

Article Snippet: Mice expressing Cre recombinase under control of D1R promoter (D1RCre) were purchased from the Jackson Laboratory (B6.FVB(129S6)-Tg(Drd1a-cre)AGsc/KndlJ strain on C57Bl/6J-congenic background).

Techniques: Injection, Immunohistochemical staining, Staining, Marker, Immunoprecipitation, Western Blot