An open-source platform for head-fixed operant and consummatory behavior
- Center for the Neurobiology of Addiction, Pain, and Emotion, Department of Anesthesiology and Pain Medicine, Department of Pharmacology, University of Washington, United States
- Department of Psychology, University of Illinois at Chicago, United States
- Graduate Program in Neuroscience, University of Illinois at Chicago, United States
Figures
Figure 1 with 1 supplement
OHRBETS (Open-Source Head-fixed Rodent Behavioral Experimental Training System) operant conditioning.
Overview of functionality for operant conditioning (additional, optional multi-solution functionality illustrated in Figure 5). (A) 3D rendering of OHRBETS. (B) Cartoon depicting the critical …
Figure 1—figure supplement 1
Validation of retractable spout and wheel brake.
(A) Cartoon of validation of the retractable spout. (B) Representative still frames from video data recording the position of the spout during 1000 extension/retractions in five different …
Figure 2 with 4 supplements
Mice rapidly learn head-fixed operant conditioning for sucrose and display operant behaviors established in freely moving experiments.
(A) Cartoon depicting the task design for retractable spout training. (B) Licking behavior throughout retractable spout training; lick raster for a representative mouse with each lick represented as …
Figure 2—figure supplement 1
Quantification of behavior during head-fixed spout training.
(A) Cartoon depicting the task design for head-fixed, free-access consumption of sucrose (free-access lick training). (B) Raster of all licks recorded in a single 10 min session for the 31 mice in …
Figure 2—figure supplement 2
Quantification of behavior during head-fixed retractable spout training.
Further quantification of behavior during retractable spout training corresponding to data shown in Figure 2G–J. (A) Lick raster of all licks recorded across all trials across three sessions for the …
Figure 2—figure supplement 3
Quantification of behavior during head-fixed operant conditioning for sucrose.
Further quantification of behavior during operant training corresponding to data shown in Figure 2L–N. (A) Cumulative position of the wheel for all mice across all sessions. (B) Density of …
Figure 2—figure supplement 4
Comparison of head-fixed and freely moving versions of operant conditioning.
(A) The total rotation of the wheel across all sessions of operant conditioning in the head-fixed version of the task (numbers on the top of the plot indicate the fixed-ratio cost of reward in wheel …
Figure 3 with 4 supplements
Head-fixed operant conditioning to obtain stimulation of lateral hypothalamic area GABAergic (LHAGABA) neurons or avoid stimulation of LHA glutamatergic (LHAGlut) neurons.
(A) Approach, placements depicted in Figure 3—figure supplement 1A. (B) Diagram of the experimental approach for positive reinforcement conducted with LHAGABA mice and negative reinforcement …
Figure 3—figure supplement 1
Placement of optic fibers.
(A) Histological locations relative to bregma of the tip of optic fibers targeting the lateral hypothalamic area (LHA). Colors indicate the experimental group of the corresponding mouse.
Figure 3—figure supplement 2
Training data for operant conditioning to obtain or avoid optogenetic stimulation.
Training data across training for each stage of the task with counts for active responses (left column), stimulations or pauses (mid column), and inactive responses (right column). (A–C) Behavior …
Figure 3—figure supplement 3
Correlation between freely moving and head-fixed stimulation count during positive reinforcement for lateral hypothalamic area GABAergic (LHAGABA) optogenetic stimulation.
(A) Correlation between the group mean z-score of stimulation counts during freely moving (abscissa) and head-fixed (ordinate) during positive reinforcement for optogenetic stimulation (error bars …
Figure 3—figure supplement 4
Optogenetic stimulation gates responding under positive and negative reinforcement.
(A) Cumulative position over a 30 min session with the laser turned off from 10 to 20 min.
Figure 4 with 3 supplements
Head-fixed wheel time preference (WTP) and aversion associated with stimulation of lateral hypothalamic area (LHA) subpopulations mirrors freely moving behavior.
(A) Approach, placements depicted in Figure 3—figure supplement 1A. (B) Cartoon depicting the freely moving and head-fixed versions of the operant task. In the head-fixed task, the mouse’s position …
Figure 4—figure supplement 1
Single subject data in the wheel time preference (WTP) and real-time place testing (RTPT) assays.
(A) Heat map showing the binned x-position in the freely moving version of the task (left three columns) or radial position in the head-fixed version of the task (right three columns) of all mice …
Figure 4—figure supplement 2
Correlation of behavior measured with the wheel time preference (WTP) and real-time place testing (RTPT) assays.
Correlation between the time spent in the paired zone during the freely moving (abscissa) or the head fixed (ordinate) at different stimulation frequencies represented as different colors for the …
Figure 4—figure supplement 3
Behavior in the wheel time preference (WTP) with and without an auditory tone.
(A) Amount of time spent in the paired zone across the freely moving (left), head fixed without a tone indicating the area the mouse was located in (middle), and head-fixed with a tone indicating …
Figure 5 with 4 supplements
Head-fixed consumption of gradients of rewarding and aversive solutions during brief access.
(A) 3D rendering of the multi-spout unit that retracts and rotates to allow brief-access periods to one of five lick spouts to the head-fixed mouse. (B) Task design. (C–I) Multi-spout consumption of …
Figure 5—figure supplement 1
Quantification of head-fixed consumption during brief access.
(A) Correlation between the number of licks and body weight gain within each session (Pearson’s product-moment correlation, r=0.87, ***p=1.15e-61). (B) Estimated volume consumed per lick produced by …
Figure 5—figure supplement 2
Multi-spout consumption of different gradients of concentrations of quinine.
Analysis of three gradients of concentrations of quinine. Each concentration set had water and four concentrations of quinine with a 1:4 serial dilution starting at 1 mM (low), 5 mM (med), and 10 mM …
Figure 5—figure supplement 3
Sex differences in head-fixed multi-spout consumption behavior.
Investigation of potential sex effects on behavior in the multi-spout brief-access assay shown in Figure 5. (A–B) Consumption of a gradient of concentrations of sucrose. (A) The mean number of licks …
Figure 5—figure supplement 4
Potential anticipation of solution identity.
Proportion of trials with a lick response in mice under water restriction during multi-spout brief-access consumption. Mice do not show of anticipation for sucrose (A, one-way RM ANOVA, …
Figure 6 with 1 supplement
Homeostatic demand shifts within session consumption of gradients of sucrose and NaCl.
(A) Procedure: Mice ran sequentially through water restriction, food restriction, and ad libitum states and during each state, mice received five sessions of multi-spout counterbalanced to have each …
Figure 6—figure supplement 1
Behavioral details for differences in consumption across homeostatic demand.
Sets of columns containing data from mice in Figure 6 undergoing multi-spout consumption of a gradient of concentrations of sucrose (left three columns) or NaCl (right two columns) across …
Figure 7 with 1 supplement
Light/dark cycle shifts within-session consumption of gradients of sucrose.
(A) Schedule for behavioral sessions. (B) Licking behavior during two sessions of free-access licking for 10% sucrose displayed as cumulative licking (left) and total lick count during the session …
Figure 7—figure supplement 1
Comparison of multi-spout behavior across labs.
(A) Data included in figure: Comparison between mice that were food-restricted and ran through multi-spout brief access to a gradient of sucrose concentrations in the dark cycle in the Stuber lab …
Figure 8 with 6 supplements
Differential dopamine dynamics during multi-spout consumption behavior.
(A) Approach for simultaneously recording dopamine dynamics in the lateral nucleus accumbens shell (NAcShL) and medial nucleus accumbens shell (NAcShM) (left), and representative placements of optic …
Figure 8—figure supplement 1
Multi-spout licking behavior.
Multi-spout licking behavior corresponding to Figure 8. (A) Mean binned lick rate for all mice for each concentration during multi-spout consumption of sucrose under food restriction (color …
Figure 8—figure supplement 2
Cumulative distribution functions of GRAB-DA responses in the NAcSh during multi-spout consumption behavior.
Cumulative distribution functions of mean GRAB-DA fluorescence during the access period for all trials with at least one lick during multi-spout sucrose under food restriction grouped and colored by …
Figure 8—figure supplement 3
Linear correlation of dopamine dynamics during multi-spout consumption.
Correlations between medial nucleus accumbens shell (NAcShM) and lateral nucleus accumbens shell (NAcShL) mean GRAB-DA fluorescence during the access period for all trials with at least one lick …
Figure 8—figure supplement 4
Range of licking and NAcSh dopamine signals during multi-spout consumption behavior.
(A) Range of licking (absolute difference in licking during access to highest and lowest concentrations) across each stage of the task (WR: water-restricted, FR: food-restricted) (one-way repeated …
Figure 8—figure supplement 5
Representative full-session traces.
(A) Full-session raw traces from mouse abb11 across three sessions of multi-NaCl under water-restricted conditions (WR:NaCl). Both the 465 nm channel (used for GRAB-DA2m imaging) and the 405 nm …
Figure 8—figure supplement 6
Fiber placements for fiber photometry.
(A) Position of fibers for fiber photometry experiments shown in Figure 8 (AP relative to bregma). Rectangles depict the fiber position determined by histology for mice included in the analysis, …
Videos
Video 1
Video of operant responding for 10% sucrose.
Video of operant responding for 10% sucrose under a fixed ratio (FR) of 1/2 turn.
Video 2
Head-fixed operant conditioning to obtain stimulation of lateral hypothalamic area GABAergic (LHAGABA) neurons or avoid stimulation of LHA glutamatergic (LHAGlut) neurons.
Videos showing responding for optogenetic stimulation of LHAGABA neurons under a positive reinforcement schedule (left) and responding for optogenetic stimulation of LHAGlut neurons under a negative …
Video 3
Consumption behavior in the multi-NaCl assay under water restriction.
Video shows licking behavior during the first 25 trials of the multi-spout assay for gradients of NaCl concentrations under water restriction. Each video depicts a single 3 s trial played back at …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | Mus musculus with name C57BL/6J | https://www.jax.org/strain/000664 | RRID:IMSR_JAX:000664 | |
| Strain, strain background (Mus musculus) | Mus musculus with name Slc32a1tm2(cre)Lowl (vgat-cre) | https://www.jax.org/strain/016962 | RRID:IMSR_JAX:016962 | |
| Strain, strain background (Mus musculus) | Mus musculus with name Slc17a6tm2(cre)Lowl (vglut2-cre) | https://www.jax.org/strain/016963 | RRID:IMSR_JAX:016963 | |
| Strain, strain background (AAV5) | AAV5-EF1a-DIO-hChR2(H134R)-eYFP | UNC Vector Core | lot #: AV4313Z | |
| Strain, strain background (AAV5) | AAV5-Ef1a-DIO-mCherry | UNC Vector Core | lot #: AV4311E | |
| Strain, strain background (AAV9) | AAV9-hSyn-GRAB-DA1h | https://www.addgene.org/113050/ | Catalog #: 113050-AAV9 | lot #: v119464 |
| Strain, strain background (AAV9) | AAV9-hSyn-GRAB-DA2m | https://www.addgene.org/140553/ | Catalog #: 140553-AAV9 | lot #: v140392 |
| Software, algorithm | Sublime Text 3 | https://www.sublimetext.com/3 | ||
| Software, algorithm | Python 3.7 (Anaconda Distribution) | https://www.anaconda.com/ | ||
| Software, algorithm | R 4.0.4 | https://cran.r-project.org/ | ||
| Software, algorithm | RStudio 2022.02.3 build 492 | https://posit.co/download/rstudio-desktop/ | ||
| Software, algorithm | Arduino IDE 1.8.13 | https://www.arduino.cc/ | ||
| Software, algorithm | TinkerCad | https://www.tinkercad.com/ | ||
| Software, algorithm | OHRBETS - Analysis v1.2; | https://github.com/agordonfennell/OHRBETS/tree/main/analysis | Author: Adam Gordon-Fennell; | |
| Other | OHRBETS - Open-source hardware | https://github.com/agordonfennell/OHRBETS | 3D printing models and bill of materials | |
| Other | Optic fiber - fiber photometry | https://www.doriclenses.com/ | MFC_400/470–0.37_6mm_MF2.5_FLT | See Materials and methods |
| Other | Optic fiber - optogenetics | https://www.rwdstco.com/ | R-FOC-BL200C-39NA | See Materials and methods; item no: 907-03007-00 |
Additional files
-
Supplementary file 1
Assembly protocol.
Detailed protocol for OHRBETS (Open-Source Head-fixed Rodent Behavioral Experimental Training System) assembly.
- https://cdn.elifesciences.org/articles/86183/elife-86183-supp1-v2.pdf
-
MDAR checklist
- https://cdn.elifesciences.org/articles/86183/elife-86183-mdarchecklist1-v2.docx
-
Source data 1
Table of statistical results.
- https://cdn.elifesciences.org/articles/86183/elife-86183-data1-v2.xlsx
