Journal: Science Advances

Article Title: A molecularly defined D1 medium spiny neuron subtype negatively regulates cocaine addiction

doi: 10.1126/sciadv.abn3552

Figure Lengend Snippet: ( A ) Viral expression of ChR2-EYFP (enhanced yellow fluorescent protein) and optic cannula placement in NAc. DAPI, 4′,6-diamidino-2-phenylindole; ac, anterior commissure. ( B ) Illustration of the two-chamber cocaine-CPP paradigm. Details are described in the “Cocaine-CPP” section. ( C ) Cocaine-CPP with optogenetic excitation of Tac2 + neurons. Left: Time spent in the cocaine-paired chambers pre- and postconditioning. Right: The CPP scores were calculated by subtracting the time spent in the preconditioning phase from the time spent in the postconditioning phase. ( D ) Cumulative distance traveled in the 30-min posttreatment period. Mice received saline or cocaine injections and were concurrently given laser stimulation [light pattern: 5 × (3 min on and 3 min off)]. ( E ) Viral expression of hM3Dq-mCherry in the NAc. ( F ) Cocaine-CPP with chemogenetic excitation of Tac2 + neurons. Left: Time spent in the cocaine-paired chambers pre- and postconditioning. Right: The CPP scores were calculated by subtracting the time spent in the preconditioning phase from the time spent in the postconditioning phase. ( G ) Viral expression of hM4Di-mCherry in the NAc. ( H ) Cocaine-CPP with chemogenetic inhibition of Tac2 + neurons. Left: Time spent in the cocaine-paired chambers pre- and postconditioning. Right: The CPP scores were calculated by subtracting the time spent in the preconditioning phase from the time spent in the postconditioning phase. Data in (C), (D), (F), and (H) are presented as means ± SEM; scale bars of (A), (E), and (G) are indicated in the figures. The P values are calculated on the basis of statistical tests in table S1. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001; ns, P > 0.05.

Article Snippet: The following AAV vectors were from Addgene: AAV5-hSyn-DIO-hM3D(Gq)-mCherry (no. 44361), AAV5-hSyn-DIO-hM4D(Gi)-mCherry (no. 44362), and AAV5-hSyn-DIO-mCherry (no. 50459).

Techniques: Expressing, Inhibition

Journal: bioRxiv

Article Title: Circuit directionality for motivation: lateral accumbens-pallidum, but not pallidum-accumbens, connections regulate motivational attraction to reward cues

doi: 10.1101/474387

Figure Lengend Snippet: Each coronal section represents a range of three sections relative to Bregma (B, in mm). Expression for each animal is plotted at 90% transparency and then per-animal expression is overlaid. Sections run anterior (top) to posterior (bottom). A) Expression map of AAV-hSyn-DIO-hM4D(Gi)-mCherry in VP following CAV2-Cre injections in NAcLSh (i.e., VP → NAcLSh inhibition animals; red). B) Expression map of AAV-hSyn-DIO-mCherry in VP (i.e., controls; black).

Article Snippet: The inhibition group received 0.6 µl of AAV-hSyn-DIO-hM4D(Gi)-mCherry (n = 12, AAV5, UNC Vector Core; n = 4, AAV8, Addgene) in the VP and 1.0 µl CAV-Cre or CAV-Cre-GFP in the NAcLSh.

Techniques: Expressing, Inhibition

Journal: bioRxiv

Article Title: Single-Cell Resolution of Individual Variation in Hypothalamic Neurons Allows Targeted Manipulation Affecting Social Motivation

doi: 10.1101/2025.03.10.642464

Figure Lengend Snippet: A. Bar plot showing the number of differentially expressed genes (DEGs) per cell type that are upregulated in Achievers (teal; positive values; n = 20) and upregulated in Non-Achievers (gray; negative values, n = 16) (*FDR adjusted p < 0.1, moderated t-test). B. Volcano plot of DEGs in PH-LHA-PVH-SO-PVa Nxph4+ glutamatergic neurons, depicting log2(fold change) versus -log2(adjusted p-value). A-enriched genes (Achiever-enriched) are shown in teal, NA-enriched (Non-Achiever-enriched) in gray, and non-significant (NS) differentially expressed genes in dark gray. Top 10 most signifcant genes are labeled. C. Dot plot showing gene ontology enrichment analysis of achiever-enriched genes (28), with significant biological processes on the y-axis. Dot size represents gene ratio, and color indicates adjusted p-value significance. D. From left to right: summary plot showing the numbers of nuclei and genes detected in each neuronal cluster (clusters shown had significantly different proportions by achiever status before FDR correction OR significant findings in regression analysis); bar plots displaying the proportions across Achiever (teal) and Non-Achiever (gray) animals for each neuronal cluster (One sample (Het_F_5) removed for plotting for STR D1/D2 Gaba cluster); and bar plot showing Beta coefficients with error bars from regression analysis measuring the relationship between cell-type-specific proportions and social rewards. * indicates significant differences at p < 0.05 after FDR corrections. E. Dot plot showing statistical significance (-log10(P-value)) of linear regression analyses for rewards as predicted by neuronal proportions, with hypothalamic regions (indigo) exhibiting marginally more significant differences than other brain regions (gray) (p = 0.08719, Fisher’s). F-G . Linear regression analyses examining relationships between PH-LHA-PVH-SO-PVa Nxph4 + glutamatergic neuron proportions and behavioral outcomes: F. Plot showing a positive relationship between mean rewards and Nxph4 + neuronal proportions, with a stronger effect in males (male: R² = 0.395; female: R² = 0.041). P-values indicate a significant effect of proportions (p = 0.009) and sex (p = 0.005). G. Plot showing no significant relationship between mean distance traveled and Nxph4+ neuronal proportions (male: R² = 0.013; female: R² = 0.023; p=0.505). H. Experimental timeline for chemogenetic inhibition studies (N = 28), showing stereotaxic injection of viral constructs (P49-57) and subsequent testing paradigm with DCZ/saline administration via voluntary oral intake of gelatin cubes during fixed ratio 1 (FR1). I. Schematic showing stereotaxic delivery of AAV5-hSyn-DIO-hM3D(Gi)-mCherry or AAV5-hSyn-DIO-mCherry control virus to the lateral and posterior hypothalamus. J. Representative confocal mosaic reconstruction of immunohistochemistry verification of viral expression in the hypothalamus, with mCherry+ cells in magenta and NeuN+ cells in teal, and DAPI staining all nuclei. K. (Left) Input-output curve showing the number of action potentials elicited after a 600ms driving current during baseline and after 1uM DCZ in mCherry+ cells in the PH (n = 5 cells from 3 animals). p values indicate a significant effect of DCZ on excitability (p = 0.0485; two-way RM ANOVA). (Right) Representative current clamp traces at 50pA (top), 100pA (middle), and 150pA (bottom) during baseline and DCZ in the same cell. L. Rheobase of mCherry+ cells during baseline and after DCZ application (p = 0.0338, paired t-test). M-O. Behavioral outcomes following chemogenetic manipulation indicate inhibition of Nxph4+ neurons in the posterior and lateral hypothalamus significantly reduces mean social rewards (p = 0.006; ANOVA), mean time in investigation zone attempting social interactions (p = 0.006; ANOVA), and overall social motivation, as measured by breakpoint, or the maximum effort exerted for a social interaction (p = 0.016; ANOVA).

Article Snippet: 360 nl of AAV5-hSyn-DIO-hM4D(Gi)-mCherry (diluted to ∼1 x 10 viral particles per mL, Addgene: 44362-AAV5) or AAV5-hSyn-DIO-mCherry (diluted to ∼1 x 10 viral particles per mL, Addgene: 50459-AAV5) was injected into PH/LHA (from bregma: ML +/-0.4 mm, AP -2.3 mm, DV - 4.9 mm).

Techniques: Labeling, Inhibition, Injection, Construct, Saline, Control, Virus, Immunohistochemistry, Expressing, Staining

Journal: Cell

Article Title: A Genetically Defined Compartmentalized Striatal Direct Pathway for Negative Reinforcement

doi: 10.1016/j.cell.2020.08.032

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-MOR Immunostar 24216 Rabbit polyclonal anti-tyrosine hydroxylase Millipore AB152 Chicken polyclonal anti-GFP Aves Labs GFP1020 Rabbit polyclonal anti-RFP Rockland 600-401-379 Rabbit monoclonal anti-HA-Tag Cell Signaling 3724S Mouse monoclonal anti-Parvalbumin Millipore MAB1572 Rabbit polyclonal anti-Somatostatin-14 Peninsula Laboratories T-4103 Goat polyclonal anti-ChAT Millipore AB144P Bacterial and Virus Strains AAV8-Ef1a-fDIO-GCaMP6m Laboratory of Karl Deisseroth N/A AAVdj-hSyn-Cre OFF /Flp ON -hChR2(H134R)-eYFP Fenno et al., 2014 Addgene 55648 AAVdj-hSyn-Cre ON /Flp OFF -hChR2(H134R)-eYFP Fenno et al., 2014 Addgene 55646 AAV8-EF1a-fDIO-Cre-p2A-mCherry Laboratory of Karl Deisseroth N/A AAV5-Ef1a-DIO-hChR2(H134R)-eYFP UNC N/A AAVdj-EF1a-fDIO-eYFP UNC N/A AAV8-hSyn-DIO-KORD-IRES-Mcitrine UNC N/A AAV2/8-Ef1a-fDIO-TVA-mCherry Laboratory of Z. Josh Huang N/A retroAAV2-CBA-fDIO-Cre Vigene Biosciences N/A AAV9-CAGGS-Flex-mKate-T2A-TVA HHMI Janelia Research Campus N/A AAV9-CAGGS-Flex-mKate-T2A-N2c-G HHMI Janelia Research Campus N/A Rbv-CVS-N2c-dG-GFP HHMI Janelia Research Campus Addgene 73461 AAV8-hSyn-DIO-mCherry Addgene Addgene 50459 Experimental Models: Organisms/Strains Mouse: Tg(Drd1a-cre)FK150Gsat/Mmucd (the “D1-Cre” line) MMRRC RRID: MMRRC_029178-UCD Mouse: B6.FVB(Cg)-Tg(Adora2a-cre) KG139Gsat/Mmucd (the “A2A-Cre” line) MMRRC RRID: MMRRC_036158-UCD Mouse: Tshz1-2A-FlpO This study N/A Mouse: Frt-Stop-Frt-TdTomato He et al., 2016 N/A Mouse: Pdyn-IRES-Cre (B6.Cg-129S-Pdyn tm1.1(cre)Mjkr/LowlJ) The Jackson Laboratory JAX: 027958 Mouse: Ai14 (B6.Cg-Gt(ROSA) 26Sortm14(CAG-tdTomato)Hze/J) The Jackson Laboratory JAX: 007908 Software and Algorithms ImageJ (Fiji) software NIH https://fiji.sc/ MATLAB Mathworks https://www.mathworks.com/ GraphPad Prism 7 GraphPad Software https://www.graphpad.com/ Open in a separate window KEY RESOURCES TABLE Tshz1 labels a population of striatal direct pathway medium spiny neurons (dMSNs) Tshz1 -expressing ( Tshz1 + ) dMSNs are localized in the striosome Tshz1 + striosomal dMSNs represent punishment and drive negative reinforcement Pdyn labels another population of striosomal dMSNs mediating positive reinforcement

Techniques: Software

Journal: Cellular and Molecular Life Sciences

Article Title: Neonatal GABAergic transmission primes vestibular gating of output for adult spatial navigation

doi: 10.1007/s00018-024-05170-x

Figure Lengend Snippet: PV-expressing VN neurons are required for navigation in adults. A 1 , B 1 Representative searching (dashed line) and returning (solid line) paths of adult (P60) mice pre-treated with saline as control group (left) or with BIC at P1 (middle) and P10 (right) under light ( A 1 ) and dark probes ( B 1 ). Filled black circles indicate the location of home base. A 2 , B 2 Graph showing the average searching time (first column), returning time (second column), heading angle (third column), and errors in locating the home base (fourth column) of these mice. * P < 0.05, ** P < 0.01, *** P < 0.001 indicate significant differences between the control and BIC-treated groups, one-way ANOVA. C 1 Schematic diagram illustrating the injection of AAV5-hSyn-DIO-hM4D Gi -mCherry into the MVN of PV-Cre mice at P42. PV-expressing MVN neurons (white arrows) with viral transfection was observed 2 weeks after injection. Scale bar, 20 μm. C 2 Trajectories of representative searching (dashed line) and returning (solid line) paths of adult PV-Cre mice expressing hM4D Gi before (left column) and after (right column) CNO administration. Paths in both light (upper panel) and dark (lower panel) probe tests are shown. C 3 Chemogenetic inhibition of PV-expressing neurons in the MVN of adult mice increased average returning time, demonstrating the importance of PV-expressing neuron activity in spatial cognitive behavior. * P < 0.05, ** P < 0.01, paired t-test. D 1 Schematic diagram illustrating the injection of control virus (AAV5-hSyn-DIO-mCherry) into the MVN of PV-Cre mice at P42. D 2 No statistical difference (paired t-test) in average searching time (first column), returning time (second column), heading angle (third column), and errors in locating the home base (fourth column) between saline and CNO treatment to these control adult mice. Data are presented as mean ± SEM

Article Snippet: Recombinant adeno-associated viruses (AAVs) carrying Cre-activated expression vectors encoding either optogenetic activator ChR2 (AAV5-hSyn-ChR2-eYFP, Penn Vector Core) or the inhibitory DREADD hM4D Gi (AAV5-hSyn-DIO-hM4D Gi -mCherry, Addgene) were purchased.

Techniques: Expressing, Saline, Control, Injection, Transfection, Inhibition, Activity Assay, Virus