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Research ArticleOpen Source Tools and Methods, Novel Tools and Methods

Custom-Built Operant Conditioning Setup for Calcium Imaging and Cognitive Testing in Freely Moving Mice

Philip Vassilev, Esmeralda Fonseca, Giovanni Hernandez, Andrea Haree Pantoja-Urban, Michel Giroux, Dominique Nouel, Elise Van Leer and Cecilia Flores
eNeuro 1 February 2022, 9 (1) ENEURO.0430-21.2022; https://doi.org/10.1523/ENEURO.0430-21.2022
Philip Vassilev
1Department of Psychiatry and Department of Neurology and Neurosurgery, McGill University, Montréal, QC, H3A 1A1, Canada
2Douglas Mental Health University Institute, Montreal, QC, H4H 1R3, Canada
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Esmeralda Fonseca
3Princeton Neuroscience Institute, Princeton University, Princeton, NJ 08540
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Giovanni Hernandez
2Douglas Mental Health University Institute, Montreal, QC, H4H 1R3, Canada
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Andrea Haree Pantoja-Urban
4Integrated Program in Neuroscience, McGill University, Montréal, QC, H3A 1A1, Canada
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Michel Giroux
2Douglas Mental Health University Institute, Montreal, QC, H4H 1R3, Canada
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Dominique Nouel
3Princeton Neuroscience Institute, Princeton University, Princeton, NJ 08540
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Elise Van Leer
2Douglas Mental Health University Institute, Montreal, QC, H4H 1R3, Canada
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Cecilia Flores
1Department of Psychiatry and Department of Neurology and Neurosurgery, McGill University, Montréal, QC, H3A 1A1, Canada
2Douglas Mental Health University Institute, Montreal, QC, H4H 1R3, Canada
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  • ORCID record for Cecilia Flores
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Figures

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  • Figure 1.
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    Figure 1.

    Overview of the setup. A desktop PC powers and controls an Arduino UNO microcontroller which in turn controls the nose poke sensors, food dispenser and audiovisual stimuli of the operant chamber. The Arduino and PC are also connected to the miniscope data acquisition box (DAQ). The DAQ, in turn, controls the USB camera for observation of the animal and the miniscope camera for recording of calcium activity. The USB camera is powered by the PC via a USB cable and the parameters of the recording can be manipulated both through the miniscope software and through other software installed for that purpose (e.g., ManyCam, Visicom Media Inc.).

  • Figure 2.
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    Figure 2.

    Operant chamber components. A–C, The base of the operant chamber is composed of the floor and the front wall with two nose poke holes and an opening for the food receptacle (A). The back of this wall holds the electronic components (B) and the pellet dispenser (C). The individual polycarbonate pieces comprising the base of the chamber are shown in (L) and their dimensions are shown in Table 1; M represents a schematic of the electronic circuit controlling the sensors and stimuli of the chamber. D–F, An additional set of removable walls limits the exploration of the mouse and focuses the behavior on the nose poke holes and pellet dispenser (optional). G–I, A removable outer casing completes the box with outer walls and two strips above the box allow for the attachment of a pipette tip box with a commutator and the USB camera (data not shown; J). The pipette box with the commutator is hinged using masking tape. K, The operant chamber in its entirety. Extended Data Figure 2-1 shows the DAQ modification for connecting to the Arduino circuit, and Extended Data Figures 2-2, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 2-11 contain the Arduino code for the Go/No-Go task.

  • Figure 3.
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    Figure 3.

    Validation of the operant setup – assessing the effect of adolescent social stress on cognition in adulthood. Adolescent mice were exposed to 4 days of accelerated social defeat stress (AcSD) and separated into resilient and susceptible groups based on a social interaction test (SIT). A, Susceptible mice had interaction ratios (IR) < 1, while resilient mice had IR >/= 1. * significantly different from control, p = 0.01. B, Susceptible mice spent less time in the interaction zone (IZ) of the SIT arena - avoiding an unfamiliar CD-1 mouse - and spent more time in the corner zones of the SIT arena (C) * significantly different from control, p = 0.02. D, The majority (60%) of mice exposed to AcSD in adolescence were resilient. E, One month after AcSD and SIT, in adulthood, mice were first trained and then tested on the Go/No-Go task. F, Schematic representation of the Go/No-Go task. Mice receive rewards by nose-poking on “Go” trials (“hits”) and withholding a nose poke during No-Go trials. Commission errors represent incorrect responses on No-Go trials. Responses before the Go (light) or No-Go (light + tone) cues on any trial are considered “premature” responses and not rewarded. G, The correct response rate represents the proportion of available rewards that were acquired by control and defeated mice across both Go and No-Go trials. H, Defeated mice made more commission errors during No-Go trials. I, There were no differences between control and defeated mice in the number of correct responses on Go trials.

  • Figure 4.
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    Figure 4.

    Calcium imaging. A, The v3 UCLA Miniscope, a representative image of mPFC neurons expressing GCaMP6f (indicated by arrows), and a representative placement of the GRIN lens. Green immunofluorescence, GcaMP6f; cyan counterstain, DAPI. Cg1, cingulate cortex; PrL, prelimbic cortex. B, A schematic representation of Go and No-Go trials depending on type of response. C, Average neuronal activity across trials centered around the presentation of both Go and No-Go cues, on days 1 and 14 of the task. Data are shown as median (±MAD) z-scored change in fluorescence (ΔF/F), n = 4. D, The same data as in C, showing Go and No-Go trials separately. E, Same data showing the neuronal response on either correct or incorrect response in the trial (Go and No-Go trials are shown together). F, The median z-scored change in fluorescence in response to Go and No-Go cues considering only significantly modulated neurons (Wilcoxon test, p < 0.0001). G, H, The same data as in F represented as change in fluorescence between 1 and −1, every row representing the fluorescence of a single neuron over time. Extended Data Figure 4-1 shows the Go/No-Go performance of the mice during the miniscope recordings.

Tables

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    Table 1

    List of polycarbonate pieces, and their dimensions, necessary to build one operant chamber (as enumerated on Fig. 2L)

    Component #DimensionsAmount (per chamber)
    111.375 × 11.375 inches1
    211.375 × 1.125 inches2
    311.875 × 3.25 inches2
    411.375 × 1.625 inches1
    52.375 × 2 inches4
    611.375 × 5.75 inches2
    79.75 × 5.875 inches2
    91.625 × 1.125 inches6
    8 and 102.375 × 1.125 inches6
    Outer enclosure18.125 × 9.75 inches2
    Outer enclosure11.5 × 9.75 inches1
    Outer enclosure12 × 1.25 inches2
    • View popup
    Table 2

    List of components necessary to build one operant chamber

    PurposeItemSupplier/product numberQuantityPrice ($ US)
    ArduinoIR break beam (for nose-poke holes)Adafruit (ID: 2167)22.33
    circuitFemale DC plugAdafruit (ID: 368)11.58
    Diode 1N4001Adafruit (ID: 755)11.19
    Jumper wiresAdafruit (ID: 760)16.28
    Speaker 2 (loud buzzer)Ajax Scientific, rectangular buzzer with lead wire, 3V13.16
    Resistor 700 ohm, 0.5WDigikey (ID: CMF55700R00FKBF-ND)10.11
    IRL 540 TransistorDigikey (ID: IRL540PBF)12.28
    Resistor 120 ohm, 0.5WMouser (ID: 588-OL1215E-R52)20.17
    Resistor 10k ohm, 0.5WMouser (ID: 603-CFR-50JR-52-10K)10.17
    White LED (very bright)Mouser (ID: 630-ASMT-AY31-NUW01)17.71
    Resistor 100 ohm, 0.5WMouser (ID: 660-CF1/2CT52A101J)20.52
    Resistor 1k ohm, 0.5WMouser (ID: 660-CF1/2CT52R102G)10.40
    28v power supply (for pellet dispenser) *Mouser (ID: 709-GST25U28-P1J) *1*19.24
    Green LEDMouser (ID: 941-C503BGCNCY0C0792)20.28
    Speaker 1 0.5W (8  Ω)SparkFun (ID: 09151)11.78
    Arduino UNO R3Sparkfun (ID: 11021)118.13
    BreadboardSparkfun (ID: 12002)13.91
    Total:*72.55
    For miniscopeCoaxial connector SMADigikey (ID: CONSMA013.062-ND)35.80
    and DAQCoaxial cable (for miniscope)Digikey (ID: A9434W-10-ND)130.22
    Coaxial cable (connections to DAQ)Digikey (ID: A9432W-10-ND)133.69
    Coaxial cable (DAQ modification)Digikey (ID: WM9477-ND)17.00
    PCB coaxial to SMAhttps://oshpark.com/shared_projects/xtQGQ32E50.59
    Total:91.26
    OtherPipette tip box, lid removedCole-Parmer (ID: RK-25712-93)130.26
    componentsCommutator *Dragonfly R&D Inc. (ID: FL-2-C-Micro)*1*346.02
    Gorilla water-resistant tapeHome Depot CA (ID: 101593)111.83
    Masking tapeHome Depot CA (ID: 2020–24)11.16
    Pellet dispenser *Med Associates*1*197.50
    USB cameraNewegg CA (ID: 9SIAVBFDST0991)118.25
    UHU White tacStaples CA (ID: 99683)13.15
    Total:*608.17
    ChamberPolycarbonate sheets (see Table 1)Local plastics storeSee Table 1134.93
    constructionSCIGRIP 16 plastics cementSciGrip Adhesives (ID: 10319)115.80
    Total:150.73
    • Optional parts and prices that include them. Please see the discussion section for alternatives.

Extended Data

  • Figures
  • Tables
  • Extended Data Figure 2-1

    Miniscope DAQ modification for external triggering of recording. A, B, A coaxial cable with an SMA connector is soldered to the input port (red square) right next to the miniscope input. C, A second coaxial cable is connected via SMA and a coaxial-to-SMA PCB. The free end is adapted to fit a breadboard by soldering two pins to the PCB. D, A top view of the operant chamber showing the miniscope DAQ (red square) connected to the miniscope via the coaxial cables (white) and the commutator. The placement of the USB camera is also visible. Download Figure 2-1, TIF file.

  • Extended Data Figure 2-2

    Arduino script for Go/No-Go training stage 0, left nose-poke hole active. Download Figure 2-2, TXT file.

  • Extended Data Figure 2-3

    Arduino script for Go/No-Go training stage 0, right nose-poke hole active. Download Figure 2-3, TXT file.

  • Extended Data Figure 2-4

    Arduino script for Go/No-Go training stage 1, left nose-poke hole active. Download Figure 2-4, TXT file.

  • Extended Data Figure 2-5

    Arduino script for Go/No-Go training stage 1, right nose-poke hole active. Download Figure 2-5, TXT file.

  • Extended Data Figure 2-6

    Arduino script for Go/No-Go training stage 2, left nose-poke hole active. Download Figure 2-6, TXT file.

  • Extended Data Figure 2-7

    Arduino script for Go/No-Go training stage 2, right nose-poke hole active. Download Figure 2-7, TXT file.

  • Extended Data Figure 2-8

    Arduino script for Go/No-Go training stage 3, left nose-poke hole active. Download Figure 2-8, TXT file.

  • Extended Data Figure 2-9

    Arduino script for Go/No-Go training stage 3, right nose-poke hole active. Download Figure 2-9, TXT file.

  • Extended Data Figure 2-10

    Arduino script for Go/No-Go training stage 4, left nose-poke hole active. Download Figure 2-10, TXT file.

  • Extended Data Figure 2-11

    Arduino script for Go/No-Go training stage 4, right nose-poke hole active. Download Figure 2-11, TXT file.

  • Extended Data Figure 4-1

    Go-No/Go performance of the 4 mice during calcium imaging. A, Overall correct response rate. B, Percent hits. C, Percent commission errors. D, Percent premature responses. Download Figure 4-1, TIF file.

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Custom-Built Operant Conditioning Setup for Calcium Imaging and Cognitive Testing in Freely Moving Mice
Philip Vassilev, Esmeralda Fonseca, Giovanni Hernandez, Andrea Haree Pantoja-Urban, Michel Giroux, Dominique Nouel, Elise Van Leer, Cecilia Flores
eNeuro 1 February 2022, 9 (1) ENEURO.0430-21.2022; DOI: 10.1523/ENEURO.0430-21.2022

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Custom-Built Operant Conditioning Setup for Calcium Imaging and Cognitive Testing in Freely Moving Mice
Philip Vassilev, Esmeralda Fonseca, Giovanni Hernandez, Andrea Haree Pantoja-Urban, Michel Giroux, Dominique Nouel, Elise Van Leer, Cecilia Flores
eNeuro 1 February 2022, 9 (1) ENEURO.0430-21.2022; DOI: 10.1523/ENEURO.0430-21.2022
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Keywords

  • animal models
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