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: Cell

Article Title: Regulation of Energy Expenditure by Brainstem GABA Neurons

doi: 10.1016/j.cell.2019.05.048

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Genotypes/sources for mice used in the above studies are listed in the . table ft1 table-wrap mode="anchored" t5 caption a7 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Chicken polyclonal anti-GFP Abcam ab13970; RRID: AB_300798 Rabbit polyclonal anti-c-Fos Synaptic Systems 226003; RRID: AB_2231974 Rabbit monoclonal anti-c-Fos Cell Signaling 2250S; RRID: AB_2247211 Rabbit polyclonal anti-RFP Rockland 600-401-379; RRID: AB_2209751 Rat monoclonal anti-mCherry (16D7) ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"M11217","term_id":"333925"}} M11217 ; RRID: AB_2536611 Chicken polyclonal anti-mCherry Abcam ab205402; RRID: AB_2722769 Chemicals, Peptides, and Recombinant Proteins Clozapine N-oxide Sigma-Aldrich C0832-5MG Critical Commercial Assays RNAscope Multiplex Fluorescent Assay V2 Advanced Cell Diagnostics (ACD Bio) 323100 RNAscope Probe- Mm-Slc32a1 ACD Bio 319191-C1 RNAscope Probe- mCherry-C2 ACD Bio 431201-C2 RNAscope Probe- EGFP ACD Bio 400281-C1 RNAscope Probe- tdTomato-C2 ACD Bio 317041-C2 Experimental Models: Organisms/Strains Mouse: Vgat-IRES-Cre: Slc32a1 tm2(cre)Lowl /J The Jackson Laboratory 016962; RRID: ISMR_JAX:016962 Mouse: Rosa26-LSL-EGFPL10a: B6.129S4- Gt(ROSA)26Sor tm1(CAG-EGFP/Rp/10a,birA)Wtp /J The Jackson Laboratory 022367; RRID: IMSR_JAX:022367 Mouse: C57BL/6J The Jackson Laboratory 000664; RRID: IMSR_JAX:000664 Mouse: TRAP2: Fos tm2.1(icre/ERT2)Luo /J The Jackson Laboratory 030323; RRID: IMSR_JAX:030323 Mouse: Ai14: B6;129S6- Gt(ROSA)26Sor tm14(CAG-tdTomato)/Hze /J The Jackson Laboratory 007908; RRID: IMSR_JAX:007908 Recombinant DNA AAV5-EF1a-DIO-hChR2(H134R)-EYFP UNC Vector Core N/A AAV5-EF1a-DIO-EYFP UNC Vector Core N/A AAV5-hSyn-DIO-hM3D(Gq)-mCherry Addgene 44361 AAV5-hSyn-DIO-hM4D(Gi)-mCherry Addgene 44362 AAV5-EF1a-fDIO-HA-NLS-Cre This paper; available at Addgene N/A AAV9-CAG-FLEX-EGFP; AI854 Penn Vector Core N/A AAV9-DIO-SynP-mCherry MGH Vector Core AAV-RN1 AAV9-DIO-SynP-EYFP MGH Vector Core AAV-RN2 PRV-lp298 (PRV-mCherry) This paper N/A PRV-152 (PRV-GFP) Smith et al., 2000 N/A Software and Algorithms GraphPad Prism 6.0 GraphPad N/A ClearMap Renier et al., 2016 N/A Imaris 9.0.1 Bitplane N/A Terastitcher University Campus Bio-Medico of Rome N/A Open in a separate window KEY RESOURCES TABLE

Techniques: Recombinant, Multiplex Assay, Fluorescence, Plasmid Preparation, Software

Journal: Nature Communications

Article Title: Human OPRM1 and murine Oprm1 promoter driven viral constructs for genetic access to μ-opioidergic cell types

doi: 10.1038/s41467-023-41407-2

Figure Lengend Snippet: a DNA sequence for the murine Oprm1 (upper) and human OPRM1 (lower) promoter regions, including the approximate locations of several transcriptional elements such as repressor and activator transcription factors (TFs), transcription start sites and promoter elements, as determined via PROMO and Eukaryotic Promoter Database & UCSC Genome Browser analyses of the murine and human genes (Supplementary Fig. ). The promoter region encoded by the four murine promoter constructs ( mMORp1-4 ) and single human promoter construct ( hMORp ) are depicted beneath each sequence map. b mMORp and hMORp construct designs and packaging schema within adeno-associated viral (AAV) vectors of multiple different capsid serotypes. c Transduction efficacy from initial in vivo intracranial injections of the mMORp1-4-eYFP constructs into C57BL/6J mouse medial prefrontal cortex (mPFC), scale bar = 500 μm. d mPFC expression pattern with AAV1- mMORp1 -eYFP across mPFC subregions, including the cingulate (Cg1), prelimbic (PL) and infralimbic (IL) cortex, scale bar = 200 μm. e Higher magnification images of the mPFC following transduction with the mMORp1 viral construct. Amplification of the eYFP signal, along with staining for both neuronal (NeuN) and microglial (Iba1) markers demonstrate selective transduction of neurons, with staining for additional glial markers to further verify this shown in Supplementary Fig. . Cortical layer division markers (Layers 1–6) highlight viral spread and efficiency, scale bar = 100 μm. f Overlap of mMORp1 -eYFP viral expression and endogenous mu opioid receptor (MOR) immunoreactivity within the central amygdala (CeA) (denoted by blue outline), but not surrounding amygdalar subregions of a C57BL/6J mouse, using a knock-out mouse-validated anti-MOR antibody (Supplementary Fig. ). Basolateral amygdala (BLA), medial anterodorsal amygdala (MeAd), medial anteroventral amygdala (MeAv), basomedial amygdala (BMA), dorsal entopeduncular nucleus (EPd), ventral entopeduncular nucleus (EPv), intercalcated cells (ITCs), globus pallidus externa (GPe), substantia innominota (SI), and lateral hypothalamus (LH); scale bar = 200 µm. g RNAscope FISH in the CeA of a mMORp1 -eYFP injected mouse examining co-localized of Oprm1 and eYfp mRNA transcripts. CeA co-localization of AAV5- mMORp1 -hM4Di-mCherry and Oprm1 2A-Cre :Sun1-sfGFP reporter nuclei ( h ) or anti-Cre staining ( i ). Co-expression of AAV5- mMORp1 -hM4Di-mCherry with anti-Cre immunoreactive cells in the CeA of Oprm1 Cre mice ( j ), and the dorsomedial striatum (DMS) ( k ), mPFC ( l ) and ventral tegmental area (VTA, m ) of Oprm1 2A-Cre mice. n Averaged number of mMORp1 -mCherry+/anti-Cre+ cells compared to mMORp1 -mCherry+/anti-Cre− cells quantified from successfully transduced brain regions of interest (from left or right hemispheres, or both) of Oprm1 Cre and Oprm1 2A-Cre :Sun1 mice within the CeA (~90.2%, N = 5, n = 9), DMS (~89.65%, N = 5, n = 7), mPFC (~88.28%, N = 3, n = 4), and VTA (~85.24%, N = 5, n = 8). Detailed information of total counts and quantification for each region within individual mouse lines can be found in Supplementary Fig. . Scale bar = 100 μm for g – m . White arrow heads denote cells in which co-labeling for Oprm1 and EYFP transcript ( g ) or mMORp1 -mCherry and anti-Cre signal ( h – m ) is observed.

Article Snippet: For cell counting studies, mice were injected with either a mix of recombinant AAV5- mMORp -hM4Di-mCherry (titer: 9.8 × 10 11 gc/mL) and AAV5- hSyn -DIO-eGFP (Addgene, #50457, titer: 1.3 × 10 13 vg/mL) at a ratio of 9ul:1ul, respectively (in Oprm1 -Cre mice) or AAV5- mMORp -hM4Di-mCherry alone (in Oprm1 2A-Cre :Sun1 mice), with similar recovery periods allowed for these animals prior to tissue collection.

Techniques: Sequencing, Construct, Transduction, In Vivo, Expressing, Amplification, Staining, Knock-Out, RNAscope, Injection, Labeling