Journal: eNeuro

Article Title: A Comprehensive, Affordable, Open-Source Hardware-Software Solution for Flexible Implementation of Complex Behaviors in Head-Fixed Mice

doi: 10.1523/ENEURO.0018-23.2023

Figure Lengend Snippet: Comparison of currently available solutions for head-fixed rodent behaviors

Article Snippet: Programming required (software base) , No# (Arduino, MATLAB) , No# (BControl, Bonsai, Alyx, Python) , No# (BControl) , OEM software** (unknown) , OEM software** (unknown) , No# (ViRMEn, MATLAB, Arduino).

Techniques: Comparison, Software

Journal: eLife

Article Title: Neural correlates and determinants of approach–avoidance conflict in the prelimbic prefrontal cortex

doi: 10.7554/elife.74950

Figure Lengend Snippet: Figure 5. In pressers, PLGLUT neurons show reduced spontaneous activity during the conflict phase. (A) Top, schematic of viral infusion. Bottom, representative immunohistochemical micrograph showing lack of colabeling (white arrows) between the ChR2 viral construct (green, AAV-CaMKII-ChR2-eYFP) and the GABA marker GAD67 (red), confirming that the use of a CaMKII promoter enables transgene expression favoring prelimbic (PL) glutamatergic neurons. Scale bars: 25 µm. (B–D) Photoidentification of PLGLUT neurons. (B) Frequency histogram

Article Snippet: DOI: https://doi.org/10.7554/eLife.74950 23 of 36 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Recombinant DNA reagent pAAV- mDlx- ChR2- mCherry- Fishell- 3 Addgene and University of North Carolina Viral Vector Core Addgene plasmid #83,898 Packaged by UNC vector core (Serotype 5) Antibody Anti- GAD67 raised in mouse polyclonal Millipore- Sigma Cat No. MAB5406B 1:400 Antibody Goat Anti- Mouse IgG H&L (Alexa Fluor 488) polyclonal Abcam Cat No. ab150113 1:200 Antibody Goat Anti- Mouse IgG H&L (Alexa Fluor 594) polyclonal Abcam Cat No. ab150116 1:200 Other Opal 520 Akoya Biosciences Cat No. FP1487001KT 1:1000 Fluorescent dye Other Opal 620 Akoya Biosciences Cat No. FP1495001KT 1:1000 Fluorescent dye Sequence- based reagent RNA scope probe mCherry Advanced Cell Diagnostics Cat No. 431201- C3 Sequence- based reagent RNA scope probe vGAT Advanced Cell Diagnostics Cat No. 424,541 Commercial Assay or Kit RNA scope Multiplex Fluorescent Detection Kit v2 Advanced Cell Diagnostics Cat No. 323,110 Chemical compound, drug VECTASHIELD Antifade Mounting Medium with DAPI Vectorlabs Cat No. H- 1200–10 Software, algorithm Matlab Mathworks RRID:SCR_001622 https://it.mathworks.com/products/matlab.html Software, algorithm NeuroExplorer Plexon RRID:SCR_001818 https://plexon.com/products/neuroexplorer/ Software, algorithm Offline Sorter Plexon RRID:SCR_000012 https://plexon.com/products/offline-sorter/ Software, algorithm CinePlex Behavioral Research System Plexon https://plexon.com/plexon-systems/cineplexbehavioral-research-system/ Software, algorithm GraphPad Prism GraphPad RRID:SCR_002798 https://www.graphpad.com/scientific-software/ prism/ Continued

Techniques: Activity Assay, Immunohistochemical staining, Construct, Marker, Expressing

Journal: eLife

Article Title: Neural correlates and determinants of approach–avoidance conflict in the prelimbic prefrontal cortex

doi: 10.7554/elife.74950

Figure Lengend Snippet: Figure 6. Photoactivation of PLGLUT, but not PLGABA, neurons suppresses reward seeking in a neutral context. (A) Schematic of viral infusion and recordings in prelimbic (PL). (B) Changes in PL firing rate with illumination of PLGLUT neurons in rats expressing AAV-CaMKII-ChR2-eYFP in PL (n = 20 neurons). (C) Average peristimulus time histogram (PSTH) of PL neurons that were excited during laser illumination of PLGLUT neurons. (D) Raster plot and PSTH of representative PL neuron showing excitatory responses to illumination in rats expressing AAV-CaMKII-ChR2-eYFP in PL. (E) Schematic of viral infusion and recordings in PL. (F) Changes in PL firing rate with illumination of PLGABA neurons in rats expressing AAV-mDlx-ChR2-mCherry in PL (n = 22 neurons). (G) Average PSTH of PL neurons that were inhibited during laser illumination of PLGABA neurons. (H) Raster plot and PSTH of representative PL neuron showing inhibitory responses to illumination in rats expressing AAV-mDlx-ChR2-mCherry in PL. Representative micrograph showing the expression of (I) CaMKII-ChR2-eYFP or (J) mDlx-ChR2-mChery in PL and schematic of optical fiber location (gray dots) in the same region (compressed across different anteroposterior levels of PL). Green or red areas represent the minimum (dark) and the maximum (light) viral expression into the PL. (K, L) Schematic and timeline of PL photoactivation during the cued food-seeking test in a neutral context. Optogenetic activation of PLGLUT neurons (CaMKII-ChR2, dark red circles, n = 7), but not PLGABA neurons (mDlx-ChR2, gold circles, n = 17), reduced the (M) frequency of lever presses (F(10, 180) = 7.009, p < 0.001, Bonferroni post hoc, CaMKII-ChR2 vs. Control, all laser on periods – p < 0.01; mDlx-ChR2 vs. Control, all laser on periods – p > 0.05) and increased (N) the latency for the first press (F(10, 180) = 9.931, p < 0.001, CaMKII-ChR2 vs. Control, all laser on periods – Bonferroni post hoc, p < 0.001; mDlx-ChR2vs. Control, all laser on periods, p > 0.05). (O) Optogenetic activation of PL neurons did not alter freezing behavior (F(10, 180) = 1.124, p = 0.346). Blue shaded area represents laser-on trials (PLGLUT: 5 Hz, PLGABA: 20 Hz; 5 ms pulse width, 7–10 mW, 30-s duration). Data shown as mean ± standard error of the mean (SEM). Each circle represents the average of two consecutive trials. Two-way repeated measures analysis of variance (ANOVA) followed by Bonferroni post hoc test. All *p’s < 0.05. All statistical analysis details are presented in Source data 1.

Article Snippet: DOI: https://doi.org/10.7554/eLife.74950 23 of 36 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Recombinant DNA reagent pAAV- mDlx- ChR2- mCherry- Fishell- 3 Addgene and University of North Carolina Viral Vector Core Addgene plasmid #83,898 Packaged by UNC vector core (Serotype 5) Antibody Anti- GAD67 raised in mouse polyclonal Millipore- Sigma Cat No. MAB5406B 1:400 Antibody Goat Anti- Mouse IgG H&L (Alexa Fluor 488) polyclonal Abcam Cat No. ab150113 1:200 Antibody Goat Anti- Mouse IgG H&L (Alexa Fluor 594) polyclonal Abcam Cat No. ab150116 1:200 Other Opal 520 Akoya Biosciences Cat No. FP1487001KT 1:1000 Fluorescent dye Other Opal 620 Akoya Biosciences Cat No. FP1495001KT 1:1000 Fluorescent dye Sequence- based reagent RNA scope probe mCherry Advanced Cell Diagnostics Cat No. 431201- C3 Sequence- based reagent RNA scope probe vGAT Advanced Cell Diagnostics Cat No. 424,541 Commercial Assay or Kit RNA scope Multiplex Fluorescent Detection Kit v2 Advanced Cell Diagnostics Cat No. 323,110 Chemical compound, drug VECTASHIELD Antifade Mounting Medium with DAPI Vectorlabs Cat No. H- 1200–10 Software, algorithm Matlab Mathworks RRID:SCR_001622 https://it.mathworks.com/products/matlab.html Software, algorithm NeuroExplorer Plexon RRID:SCR_001818 https://plexon.com/products/neuroexplorer/ Software, algorithm Offline Sorter Plexon RRID:SCR_000012 https://plexon.com/products/offline-sorter/ Software, algorithm CinePlex Behavioral Research System Plexon https://plexon.com/plexon-systems/cineplexbehavioral-research-system/ Software, algorithm GraphPad Prism GraphPad RRID:SCR_002798 https://www.graphpad.com/scientific-software/ prism/ Continued

Techniques: Expressing, Activation Assay, Control