Journal: Cell

Article Title: Endogenous opioid dynamics in the dorsal striatum sculpt neural activity to promote goal-directed action

doi: 10.1016/j.cell.2026.07.058

Figure Lengend Snippet: ( A ) Schematic of viral injection in the DMS and operant behavior. ( B ) Confocal images of Ctrl (top) and DMS pdyn-cKO (bottom) with ISH for DAPI (blue) and DRD1 (green), and DAPI (blue) and Pdyn (majenta) for sections of Striatum (left, 10X), DMS (middle, 40X) and Nac (right), and Quantification of ISH. ( C ) Operant learning (Simple Linear Regression: n=9 Ctrl mice, R 2 =0.5782, p<0.0001**** . n=12 DMS pdyn-cKO mice, R 2 =0.2808, p=0.0009*** . Difference in Slopes – p=0.0229* ). ( D ) Operant Reversal (Simple Linear Regression: n=5 Ctrl mice, R 2 =0.7663, p<0.0001**** . n=5 DMS pdyn-cKO mice, R 2 =0.7333, p<0.0001**** . Difference in Slopes – p=0.0133* ). ( E ) Operant FR-3 (Simple Linear Regression: n=5 Ctrl mice, R 2 =0.2814, p=0.0419* . n=5 DMS pdyn-cKO mice, R 2 =0.04635, p>0.05 . Difference in Intercepts – p=0.0029 ## ). ( F ) Operant Extinction (Simple Linear Regression: n=5 Ctrl mice, R 2 =0.7928, p<0.0001**** . n=5 DMS pdyn-cKO mice, R 2 =0.1324, p>0.05 . Difference in Slopes – p=0.0086** ). ( G ) Operant U50,488 injection (n=5 Ctrl, DMS pdyn-cKO mice; Two Way ANOVA, genotype x treatment p<0.0037** . Multiple comparisons – Ctrl veh vs. U50, p>0.05; DMS pdyn-cKO veh vs. U50, p=0.0003*** ). ( H ) Schematic of viral injection and fiber implant in the DMS. ( I ) Left – Learning (Simple Linear Regression: n=6 Ctrl mice, R 2 =0.8036, p<0.0001**** . n=5 DMS pdyn-cKO mice, R 2 =0.3523, p=0.0197* . Difference in Slopes – p=0.0003*** ). Right - Extinction (Simple Linear Regression: n=6 Ctrl mice, R 2 =0.7947, p<0.0001**** . n=5 DMS pdyn-cKO mice, R 2 =0.2682, p=0.0480* . Difference in Slopes – p=0.0011** ) ( J-L ) Mean fluorescence and heatmap raster plots during early, trained and extinction operant behavior (representative animals) for Ctrl (dark) and DMS pdyn-cKO (light). ( M ) Normalized Peak z-score values during cue+outcome (2–10s) of Ctrl (Left: n=6 mice; One Way ANOVA, p=0.0004 ***. Multiple comparisons – early vs. trained, p<0.0001**** , trained vs. extinction, p=0.0029** ) and DMS pdyn-cKO (n=5 mice; One Way ANOVA, p>0.05 ). ( N ) Peak z-score values during cue+outcome (2–10s) subtracted from a baseline (−5 to −1s) (n=6 Ctrl, 5 DMS pdyn-cKO mice; Two Way ANOVA, genotype x day p<0.0037** . Multiple comparisons – Early, p>0.05; Trained, p<0.0001**** ; Extinction, p=0.0359* ).

Article Snippet: VECTASHIELD Hardset Antifade Mounting Medium with DAPI , Vector Laboratories , CAT#H-1800.

Techniques: Injection, Confocal, Fluorescence

Journal: Cell

Article Title: Endogenous opioid dynamics in the dorsal striatum sculpt neural activity to promote goal-directed action

doi: 10.1016/j.cell.2026.07.058

Figure Lengend Snippet: ( A ) Top - Schematic of viral injection and optic fiber implantation in the DMS. Bottom - 20X Confocal image of optic fiber implant in the DMS with DAPI, expressing κLight 1.3a. ( B ) Top - Operant behavior schedule during photometry. Bottom: Operant behavior (n=6 mice, Left, Learning Simple Linear Regression: R 2 =0.8382, p<0.0001 . Right, Extinction Simple Linear Regression: R 2 =0.5800, p=0.0002 ). ( C-E ) Mean fluorescence and heatmap raster plots during early, trained and extinction operant behavior (representative animal). ( F ) Normalized Peak z-score values during cue+outcome (2–10s) (n=6mice; One Way ANOVA, p<0.0001 ****. Multiple comparisons – early vs. trained, p=0.0307* , trained vs. extinction, p=0.0006*** , early vs. extinction, p=0.0094** ). ( G ) Schematic of Generalized Linear Model (GLM) analyses for trial-trial peak z-score values during cue+outcome and behavioral variables – trial number ( trial, green ), preceding inter-trial interval ( ITI, light purple ), subsequent inter-trial interval ( ITI+1, dark purple ) and latency to consume the reward pellet following a reinforced active nosepoke ( eat, pink ). ( H ) Proportion of variance following behavioral variable drop-out in the trained sessions (n=6 mice, 199 trials; Full Model R 2 =0.4414, Trial R 2 =0.4755, p>0.05 , ITI R 2 =0.4724, p>0.05 , ITI+1 R 2 =0.0751, p=0.047* , Eat R 2 =0.4257, p>0.05 ). ( I ) Mean Fluorescence for three consecutive trials with short ITIs (top) and two consecutive trials with long ITIs (bottom) time-locked to active nosepokes from a representative animal. ( J ) Heat maps for trial-trial normalized ITIs (left) and trial-trial peak z-score values during cue+outcome (right) from a representative animal. ( K ) Correlation between normalized ITI+1 and peak z-score during cue+outcome (Second Order Polynomial Quadratic Fit, R 2 =0.5408, p<0.0001**** ). ( L ) Top - Schematic of viral injection and optic fiber implantation in the DMS. Bottom - 20X Confocal image of optic fiber implant in the DMS with DAPI, expressing ChR2. ( M ) Operant behavior during photoactivation (n=8 Ctrl,8 dyn-cre mice; Two Way ANOVA, stim x genotype p<0.0009*** . Multiple comparisons – ctrl off vs. on, p>0.05; dyn-cre off vs. on, p<0.0001**** ). ( N ) Inter-trial Intervals for operant behavior during photoactivation (n=8 Ctrl,8 dyn-cre mice; Two Way ANOVA, stim x genotype p=0.0005*** . Multiple comparisons – ctrl off vs. on, p>0.05; dyn-cre off vs. on, p=0.0005*** ). ( O ) Extinction during days 1–5 (Left, Simple Linear Regression: n=4mice, Off - R 2 =0.6401, p=0.0018** , On – R 2 =0.001827, p>0.05 . Difference in intercepts – p=0.0035 ## ), and days 6–10 (Left, Simple Linear Regression: n=4mice, Off - R 2 =0.6401, p<0.0001**** , On – Simple Linear Regression, R 2 =0.008760, p>0.05 . Difference in Slopes – p=0.0356* ). ( P ) Operant behavior during photoactivation with aticaprant injection (n=8 dyn-cre mice; Two Way ANOVA, stim x treatment p<0.0059** . Multiple comparisons – veh off vs. on, p=0.0007***; aticaprant off vs. on, p>0.05 ).

Article Snippet: VECTASHIELD Hardset Antifade Mounting Medium with DAPI , Vector Laboratories , CAT#H-1800.

Techniques: Injection, Confocal, Expressing, Fluorescence

Journal: Cell

Article Title: Endogenous opioid dynamics in the dorsal striatum sculpt neural activity to promote goal-directed action

doi: 10.1016/j.cell.2026.07.058

Figure Lengend Snippet: ( A ) Top - Schematic of viral injection and prism implantation in the DMS. Bottom – 20X and 40X confocal images of prism implant in the DMS with DAPI, expressing GCaMP6s and Td-Tomato. ( B ) Top – Representative standard deviation images of GCaMP6s (left) and Td-tomato (right) fluorescence. Bottom – tracked and co-registered GCaMP6s+Td-tomato neurons and proportion of tracked GCaMP6s+ Td-tomato neurons across 3 mice. ( C ) Top - Schematic of experiment and operant behavior schedule using a head-fixed wheel-based task. Bottom - Operant Behavior (n=3mice, Simple Linear Regression: Learning - R 2 =0.7388, p=0.003** , Extinction - R 2 =0.9837, p=0.0001**** ). ( D-F ) Spectral clustering classification of early, trained and extinction operant data across 3 mice showing fluorescence activity traces (top) and heat maps (bottom) of cells. ( G ) Peak z-score quantification of clusters - Action (549 cells; One Way ANOVA, p<0.0041** . Multiple comparisons – trained vs. early, p<0.0122* , trained vs. extinction, p<0.0030** ), Action+Cue (487 cells; One Way ANOVA, p<0.0196* . Multiple comparisons – trained vs. early, p=0.0344* , trained vs. extinction, p=0.0169* ), Cue+Reward clusters (332 cells; One Way ANOVA, p<0.0485* . Multiple comparisons – trained vs. early, p>0.05 , trained vs. extinction, p=0.0344* ), Reward (345 cells; One Way ANOVA, p<0.0302* . Multiple comparisons – trained vs. early, p>0.05 , trained vs. extinction, p=0.0239* ), and Inactive (967 cells; One Way ANOVA, p=0.0153* . Multiple comparisons – trained vs. early, p=0.01* , trained vs. extinction, p>0.05 ). ( H ) Schematic of Artificial Recurrent Neural Network (RNN) modelling for decoding trial-trial DMS pdyn neuron activity to predict ITI length split into quintiles based on length using the activity of the preceding trial (trial N; ITI+1), subsequent trial (trial N+1; ITI), or both. ( I ) Accuracy of RNN model for early (n=3 mice, 38 trials, 2680 cells; shuffled vs. real neural data, unpaired t test, p>0.05 ), trained (n=3 mice, 182 trials, 2680 cells; shuffled vs. real neural data, unpaired t test, p<0.0001**** ) and extinction (n=3 mice, 70 trials, 2680 cells; shuffled vs. real neural data, unpaired t test, p>0.05 ). ( J ) Accuracy of RNN model for shortest 1/5 th ITIs (Left, n=3 mice, 37 trials, 2680 cells; One Way ANOVA, p<0.0001**** . Multiple comparisons – trained vs. early, p=0.0103* , trained vs. extinction, p=0.0002*** ) and longest 1/5 th ITIs (Right, n=3 mice, 37 trials, 2680 cells; One Way ANOVA, p>0.05 ). ( K ) Schematic of identifying neurons belonging to specific clusters based on the accuracy of decoding ITI length using the RNN model. ( L ) Accuracy of RNN model after dropping out specific clusters for shortest one-fifth of ITIs length in trained operant (n=3 mice, 37 trials; Dashed Line – Full Model Accuracy, One Way ANOVA, p=0.0030** . Multiple comparisons – full vs. action, p>0.05 , full vs. action+cue, p=0.0268* , full vs. cue+reward, p>0.05, full vs. reward, p>0.05 , full vs. inactive, p>0.05 ). ( M ) Schematic describing hypothesis: DMS pdyn neuron activity and dynorphin release evolves as animals learn goal-directed behavior, segregating their activity in response to distinct behavioral variables (left). Upon learning, DMS pdyn neuron activity and dynorphin release influences engagement in goal-directed action-outcome behavior (right).

Article Snippet: VECTASHIELD Hardset Antifade Mounting Medium with DAPI , Vector Laboratories , CAT#H-1800.

Techniques: Activity Assay, Injection, Confocal, Expressing, Standard Deviation, Fluorescence

Journal: Cell

Article Title: Endogenous opioid dynamics in the dorsal striatum sculpt neural activity to promote goal-directed action

doi: 10.1016/j.cell.2026.07.058

Figure Lengend Snippet: ( A ) Top - Schematic of viral injection in the DMS. Middle - 20X (left) and 40X (middle) Confocal image of BLA section with ISH for DAPI, CamKII, KOR and Cre, and (right) Quantification of ISH. Bottom - 20X (left) and 40X (middle) Confocal image of BLA section with ISH for DAPI, CamKII, Vglut1 and Cre, and (right) Quantification of ISH. ( B ) Top - Schematic of viral injection in the BLA. 20X (middle) and 40X (bottom) confocal image of DMS sections stained for DAPI, with Vglut1 + (left) or KOR + (right) BLA fibers expressing ChR2. ( C ) Schematic of viral injection in the BLA. ( D ) Input-output curve of optically-evoked EPSCs from DMS pdyn and D1(−) DMS neurons (n=5 mice, 16 cells; Two Way ANOVA, Cell-type p<0.0014** . Multiple comparisons – DMS pdyn vs. D1(−) at 2.7 mW, p=0.0240 , at 5.7 mW p=0.0282* ). ( E ) Normalized optically-evoked EPSCs from DMS pdyn and D1(−) DMS neurons following U69 (n=4 mice, 8 cells; DMS pdyn - paired t test, p<0.0313* ; D1(−) - paired t test, p<0.0002*** ). ( F ) Normalized optically-evoked EPSCs from DMS neurons following U69 (n=5 mice, 17 cells, D1-TdTom; n=4 mice, 6 cells, KOR-cKO; D1-TdTom vs. KOR-cKO - unpaired t test, p<0.0235* ). ( G ) Representative traces for Veh and U69 in DMS (red) and KOR-cKO (brown). ( H ) Left - Schematic of viral injection in the BLA and schematic of operant behavior. Right - 20X (left) and 40X (middle) Confocal image of Control (top) and BLA KOR-cKO (bottom) BLA section with ISH for DAPI, CamKII, KOR and overlap, and (right) Quantification of ISH. ( I ) Left – Learning (Simple Linear Regression: n=6 Ctrl mice, R 2 =0.6587, p<0.0001**** . n=8 BLA KOR-cKO mice, R 2 =0.2152, p=0.0224* . Difference in Slopes – p=0.0043** ). Right – Extinction (Simple Linear Regression: n=6 Ctrl mice, R 2 =0.5057, p=0.0009*** . n=8 BLA KOR-cKO mice, R 2 =0.0440, p>0.05 . Difference in Slopes – p=0.001** ).

Article Snippet: VECTASHIELD Hardset Antifade Mounting Medium with DAPI , Vector Laboratories , CAT#H-1800.

Techniques: Injection, Confocal, Staining, Expressing, Control

Journal: Cell

Article Title: Endogenous opioid dynamics in the dorsal striatum sculpt neural activity to promote goal-directed action

doi: 10.1016/j.cell.2026.07.058

Figure Lengend Snippet: ( A ) Top - Schematic of viral injection in the BLA and optic fiber implantation in the DMS. Bottom - 20X Confocal image of optic fiber implant in the DMS and BLA with DAPI, expressing GCaMP6s. ( B ) Top - Operant behavior schedule during photometry. Bottom – Operant (n=6 mice; Simple Linear Regression: Learning, R 2 =0.7781, p<0.0001**** . Extinction, R 2 =0.4251, p=0.0033** ). ( C-E ) Mean fluorescence and heatmap raster plots during early, trained and extinction operant behavior from a representative animal. ( F ) Top – Normalized Peak z-score values during action (−20–0s, top) (n=6 mice; One Way ANOVA, p=0.0026 **. Multiple comparisons – early vs. trained, p=0.0005*** , trained vs. extinction, p=0.0092** , early vs. extinction, p>0.05 ), and cue+outcome (2–10s, bottom) (n=6 mice; p=0.0002 *. Multiple comparisons – early vs. trained, p=0.0002*** , trained vs. extinction, p=0.0027* , early vs. extinction, p>0.05 ). ( G ) Simple Linear Regression between normalized ITI vs. peak z-score during action across early (n=6 mice, 78 trials, R 2 =0.002, p>0.05 ), trained (n=6 mice, 290 trials, R 2 =0.4733, p<0.0001**** ) and extinction (n=6mice, 526 trials, R 2 =0.0055, p>0.05 ). ( H ) Simple Linear Regression between normalized ITI vs. peak z-score during cue+outcome across early (n=6 mice, 78 trials, R 2 =0.00525, p>0.05 ), trained (n=6mice, 290 trials, R 2 =0.0024, p>0.05 ) and extinction sessions (n=6mice, 526 trials, R 2 =0.0014, p>0.05 ). ( I ) Simple Linear Regression between normalized ITI+1 vs. peak z-score during action across early (n=6 mice, 72 trials, R 2 =0.0209, p>0.05 ), trained (n=6 mice, 284 trials, R 2 =0.0204, p>0.05 ) and extinction (n=6mice, 520 trials, R 2 =0.0016, p>0.05 ). ( J ) Simple Linear Regression between normalized ITI+1 vs. peak z-score during action across early (n=6mice, 72 trials, R 2 =0.007, p>0.05 ), trained (n=6mice, 284 trials, R 2 =0.3744, p<0.0001**** ) and extinction sessions (n=6mice, 520 trials, R 2 =0.0003, p>0.05 ). ( K ) Left - Schematic of viral injection in the BLA and optic fiber implantation in the DMS. Right - 20X Confocal image of optic fiber implant in the DMS (top) and BLA (bottom) with DAPI, expressing ChR2. ( L ) Active nosepokes for 1 second self-stimulation (n=8 vglut1-cre mice; paired t test, p<0.0001**** ). ( M ) Operant behavior during photoactivation at action (n=4 Ctrl,8 vglut1-cre mice; Two Way ANOVA, stim x genotype p>0.05 ). ( N ) Operant behavior during photoactivation at cue+outcome (n=4 Ctrl,8 Vglut1-cre mice; Two Way ANOVA, stim x genotype p=0.005** . Multiple comparisons – ctrl off vs. on, p>0.05; vglut1-cre off vs. on, p=0.0008*** ). ( O ) Inter-trial Intervals for operant behavior during photoactivation at cue+outcome (n=4 Ctrl,8 Vglut1-cre mice; Two Way ANOVA, stim x genotype p=0.0041** . Multiple comparisons – ctrl off vs. on, p>0.05; vglut1-cre off vs. on, p<0.0001**** ). ( P ) Left - Schematic of viral injection in the BLA and optic fiber implantation in the DMS. Right - 20X Confocal image of optic fiber implant in the DMS (top) and BLA (bottom) with DAPI, expressing PPO. ( Q ) Operant behavior during photoinhibition at whole session (n=4 Ctrl,9 vglut1-cre mice; Two Way ANOVA, stim x genotype p=0.0079** . Multiple comparisons – ctrl off vs. on, p>0.05; vglut1-cre off vs. on, p<0.0001**** ). ( R ) Operant behavior during photoinhibition at cue+outcome (n=4 Ctrl,9 vglut1-cre mice; Two Way ANOVA, stim p=0.0005*** , stim x genotype p>0.05 . Multiple comparisons – ctrl off vs. on, p>0.05; vglut1-cre off vs. on, p=0.0002*** ). ( S ) Inter-trial Intervals for operant behavior during photoinhibition at cue+outcome (n=4 Ctrl,9 Vglut1-cre mice; Two Way ANOVA, stim x genotype p=0.005*** . Multiple comparisons – ctrl off vs. on, p>0.05; vglut1-cre off vs. on, p<0.0001**** ). ( T ) Extinction behavior during photoinhibition at whole session (Simple Linear Regression: n=5 vglut1-cre mice; Off - R 2 =0.6151, p=0.0005*** . On – R 2 =0.2822, p=0.0416* . Difference in Slopes – p=0.0261* ).

Article Snippet: VECTASHIELD Hardset Antifade Mounting Medium with DAPI , Vector Laboratories , CAT#H-1800.

Techniques: Injection, Confocal, Expressing, Fluorescence

Journal: Cell

Article Title: Endogenous opioid dynamics in the dorsal striatum sculpt neural activity to promote goal-directed action

doi: 10.1016/j.cell.2026.07.058

Figure Lengend Snippet: ( A ) Top - Schematic of viral injection in the BLA and optic fiber implantation in the DMS. Bottom: 10X (left) and 40X (right) confocal images of Vglut1-Cre sgROSA (top) and Vglut1-Cre oprk1 (bottom) BLA section with ISH for DAPI (blue) and Vglut1 (green), and DAPI (blue) and KOR (majenta), and quantification of ISH. ( B ) Operant learning (Simple Linear Regression: n=4 Vglut1-Cre sgROSA mice, R 2 =0.9173, p<0.0001**** . n=6 Vglut1-Cre oprk1 mice, R 2 =0.6745, p<0.0001**** . Difference in Slopes – p<0.0001**** ) ( C ) Mean fluorescence and heatmap raster plots during operant behavior (n=4 Vglut1-Cre sgROSA , 6 Vglut1-Cre oprk1 mice). ( D ) Peak z-score values during action (−5–0s), subtracted from baseline (−10 to −5s) (n=4 Vglut1-Cre sgROSA , 6 Vglut1-Cre oprk1 mice; unpaired t test, p=0.0042** ). ( E ) Minimum z-score values during cue+outcome (2–10s), subtracted from baseline (−10 to −5s) (n=4 Vglut1-Cre sgROSA , 6 Vglut1-Cre oprk1 mice; unpaired t test, p=0.0483* ). ( F ) Left - Schematic of viral injection in the BLA and DMS, and optic fiber implantation in the DMS. Right - 20X Confocal image of optic fiber implant in the DMS (top) and BLA (bottom) with DAPI, expressing GCaMP6s. ( G ) Operant learning (Simple Linear Regression: n=4 Ctrl mice, R 2 =0.8551, p<0.0001**** . n=7 DMS pdyn-cKO mice, R 2 =0.3656, p=0.0037** . Difference in Slopes – p<0.0001**** ). ( H ) Mean fluorescence and heatmap raster plots during operant behavior (n=4 Ctrl mice, n=7 DMS pdyn-cKO mice). ( I ) Peak z-score values during action (−5–0s), subtracted from baseline (−10 to −5s) (n=4 Ctrl, 7 DMS pdyn-cKO mice; unpaired t test, p=0.0001*** ). ( J ) Minimum z-score values during cue+outcome (2–10s), subtracted from baseline (−10 to −5s) (n=4 Ctrl, 7 DMS pdyn-cKO mice; unpaired t test, p=0.0497* ). ( K ) Schematic of viral injection in the BLA and DMS, and optic fiber implantation in the DMS. ( L ) 10X (left) and 40X (right) confocal images of DMS TScKO Striatum section with ISH for DAPI (blue) and DRD1 (green), DAPI (blue) and Pdyn (majenta), and DAPI (blue) and Cre (gold). Bottom - Quantification of ISH. ( M ) Operant learning (Simple Linear Regression: n=4 Ctrl mice, R 2 =0.4692, p=0.014* . n=6 DMS TScKO mice, R 2 =0.6239, p<0.0001**** . Difference in Slopes – p=0.0204* ).

Article Snippet: VECTASHIELD Hardset Antifade Mounting Medium with DAPI , Vector Laboratories , CAT#H-1800.

Techniques: Injection, Confocal, Fluorescence, Expressing