A virtual burrow assay for head–fixed mice measures habituation, discrimination, exploration and avoidance without training
- Howard Hughes Medical Institute, Columbia University, United States
Figures
Figure 1 with 4 supplements
The Virtual Burrow Assay.
(A) Left; diagram of ingress (top) and egress (bottom). Right, photograph of mouse in ingress position (top) and egress position (bottom). (B) Instrument diagram. The mouse’s head is fixed by a …
Figure 1—figure supplement 1
Persistence of stereotyped behavior across multiple days.
(A) Distribution of tube position during open loop mode on four separate days over a 16 day interval for the same cohort of N = 5 mice. We used a Wilcoxon rank-sum test to evaluate whether burrow …
Figure 1—figure supplement 2
Schematic and flow diagram.
(A) Schematic of the virtual burrow control system. Control inputs and outputs, black. Devices, gray. (B) Finite state machine diagram. During the intertrial interval (ITI), a linear actuator …
Figure 1—video 1
Spontaneous behavior of a mouse in the Virtual Burrow Assay.
Video recorded immediately following the mouse’s first introduction into the Virtual Burrow Assay.
Figure 1—video 2
Burrow pulled to egress position.
Left: video of the same mouse shown in Figure 1—video 1 taken approximately two minutes later during the first instances in which it was pulled out to the egress position when the command software …
Figure 2 with 1 supplement
Reliable, short-latency ingress to noxious air puff.
(A) Diagram of experimental set up. The mouse is head-fixed in the virtual burrow and an air puff is delivered to the nose. (B) Left, burrow position over time showing a single ingress in response …
Figure 2—video 1
Ingress in response to strong air puff.
White square: stimulus epoch (250 msec, 80 psi, 2 mm distance from the snout).
Figure 3 with 1 supplement
Flight-inducing visual stimuli selectively elicit ingress.
(A) An expanding black disk (left, orange), a contracting black disk (middle, pink), and a sweeping black disk of constant size (right, green), were presented on a visual display positioned directly …
Figure 3—video 1
Ingress in response to visual looming stimulus.
Top left panel: visual stimulus presented on a screen positioned above the animal’s head (see diagram in Figure 3A); bottom panel: video of a mouse head-fixed in the VBA; top right panel: position …
Figure 4 with 1 supplement
Habituation to unanticipated stimuli.
(A) Three odorant stimuli were delivered to mice in the virtual burrow assay. (B) Habituating responses to repeated presentation of odorant stimuli from a representative mouse. Color map corresponds …
Figure 4—video 1
Egress in response to unanticipated odorant stimulus.
Responses to the first two presentations of an odorant stimulus. White square: stimulus epoch (8 s).
Figure 5 with 2 supplements
Ingress in response to aversively conditioned odorant stimuli.
(A) Three odorant stimuli were presented to mice head fixed in the VBA on day 1 (Pre-test) and day 3 (Test). On day 2 (Conditioning), animals were placed in a fear conditioning chamber and two of …
Figure 5—figure supplement 1
Trembling in response to aversively conditioned odorant stimuli.
Tube position during three individual trials recorded during Test in response to CS + and CS-. Odorant stimulus epoch denoted by the black bar (top); dashed box at left demarcates epoch in which …
Figure 5—figure supplement 2
Robustness of statistical test to choice of ingress threshold.
Effect of maximum displacement threshold on the two-proportion z-test p value for the pooled test data (nine mice, six trials per mouse). Dashed line indicates p=0.05.
Figure 6
Latency of ingress to aversively conditioned odorant stimuli.
(A) Distribution of ingress onset latency during CS+ trials for all mice. Ingress onset defined as the first sample in which displacement exceeded 0.75 mm following stimulus onset. (B) Ingress onset …
Figure 7
Simultaneous measurement of behavior and neuronal activity.
(A) Top to bottom: position, velocity, force, mean firing rate and raster plot of 133 simultaneously recorded single-units in anterior piriform cortex of an awake mouse in the VBA. Left; 2 min epoch …
Tables
Table 1
Animals used in this study.
Identifying information for all animals used in this study.
| Animal number | Age at surgery | Age at expt. | Data | Notes |
|---|---|---|---|---|
| 2017022711 | 17 wks | 31 wks | Ingress, Figure 1C,D | - |
| 2017022712 | 17 wks | 31 wks | Ingress, Figure 1C,D | Also used for air puff expt. |
| 2017031302 | 10 wks | 22 wks | Ingress, Figure 1C,D | Also used for air puff expt. |
| 2017032101 | 11 wks | 22 wks | Ingress, Figure 1C,D | - |
| 2017032401 | 8 wks | 19 wks | Ingress, Figure 1C,D | - |
| 2017032402 | 11 wks | 22 wks | Ingress, Figure 1C,D | - |
| 2017033001 | 9 wks | 20 wks | Ingress, Figure 1C,D | Also used for air puff expt. |
| 2017033002 | 9 wks | 20 wks | Ingress, Figure 1C,D | Also used for air puff expt. |
| 2017042103 | 12 wks | 20 wks | Ingress, Figure 1C,D | - |
| 2017042105 | 12 wks | 20 wks | Ingress, Figure 1C,D | Also used for air puff expt. |
| 2018032602 | 8 wks | 14–16 wks | Figure 1—figure supplement 1 | |
| 2018040204 | 9 wks | 15–17 wks | Figure 1—figure supplement 1 | |
| 2018040206 | 9 wks | 15–17 wks | Figure 1—figure supplement 1 | |
| 2018040208 | 9 wks | 15–17 wks | Figure 1—figure supplement 1 | |
| 2018040212 | 9 wks | 15–17 wks | Figure 1—figure supplement 1 | |
| 2016082207 | 11 wks | 18 wks | Air puff, Figure 2 | Figure 2B,C from this animal |
| 2017022712 | 17 wks | 31 wks | Air puff, Figure 2 | Also used for Figure 1C,D |
| 2017031302 | 10 wks | 22 wks | Air puff, Figure 2 | Also used for Figure 1C,D |
| 2017033001 | 9 wks | 20 wks | Air puff, Figure 2 | Also used for Figure 1C,D |
| 2017033002 | 9 wks | 20 wks | Air puff, Figure 2 | Also used for Figure 1C,D |
| 2017042105 | 12 wks | 20 wks | Air puff, Figure 2 | Also used for Figure 1C,D |
| 2018061501 | 7 wks | 15 wks | Air puff, Figure 2 | - |
| 2018061201 | 7 wks | 15 wks | Air puff, Figure 2 | - |
| 2018061202 | 7 wks | 15 wks | Air puff, Figure 2 | - |
| 2018061301 | 7 wks | 15 wks | Air puff, Figure 2 | - |
| 2016071401 | 15 wks | 21 wks | Visual stimuli, Figure 3 | Expanding disk |
| 2016072401 | 16 wks | 21 wks | Visual stimuli, Figure 3 | Expanding disk |
| 2016072702 | 17 wks | 21 wks | Visual stimuli, Figure 3 | Expanding disk |
| 2016081901 | 10 wks | 11 wks | Visual stimuli, Figure 3 | Receding disk |
| 2016081902 | 10 wks | 11 wks | Visual stimuli, Figure 3 | Receding disk |
| 2016081903 | 10 wks | 11 wks | Visual stimuli, Figure 3 | Receding disk |
| 2015111301 | 15 wks | 56 wks | Visual stimuli, Figure 3 | Sweeping disk |
| 2015111602 | 12 wks | 52 wks | Visual stimuli, Figure 3 | Sweeping disk |
| 2015111701 | 12 wks | 52 wks | Visual stimuli, Figure 3 | Sweeping disk |
| 2016082302 | 11 wks | 18 wks | Odor habituation, Figure 4A–D | O1: Lim. O2: Oct. O3: Hex. |
| 2016082401 | 14 wks | 21 wks | Odor habituation, Figure 4A–D | O1: Oct. O2: Hex. O3: Lim. |
| 2016082402 | 14 wks | 21 wks | Odor habituation, Figure 4A–D | O1: Hex. O2: Oct. O3: Lim. |
| 2016082404 | 14 wks | 21 wks | Odor habituation, Figure 4A–D | O1: Hex. O2: Oct. O3: Lim |
| 2016082405 | 14 wks | 21 wks | Odor habituation, Figure 4A–D | O1: Oct. O2: Hex. O3: Lim. |
| 2017071603 | 13 wks | 34 wks | Odor habituation, Figure 4E–H | - |
| 2017071401 | 13 wks | 34 wks | Odor habituation, Figure 4E–H | - |
| 2017071605 | 13 wks | 34 wks | Odor habituation, Figure 4E–H | - |
| 2017071601 | 13 wks | 34 wks | Odor habituation, Figure 4E–H | - |
| 2017071602 | 13 wks | 34 wks | Odor habituation, Figure 4E–H | - |
| 2016082006 | 11 wks | 21 wks | Aversive odor learning, Figure 5, 6 | - |
| 2016082005 | 11 wks | 21 wks | Aversive odor learning, Figure 5, 6 | - |
| 2016082003 | 11 wks | 21 wks | Aversive odor learning, Figure 5, 6 | - |
| 2016082001 | 11 wks | 21 wks | Aversive odor learning, Figure 5, 6 | - |
| 2016071401 | 15 wks | 30 wks | Aversive odor learning, Figure 5, 6 | Also used for Figure 3 |
| 2016081903 | 10 wks | 20 wks | Aversive odor learning, Figure 5, 6 | Also used for Figure 3 |
| 2016081902 | 10 wks | 20 wks | Aversive odor learning, Figure 5, 6 | Also used for Figure 3 |
| 2016081901 | 10 wks | 20 wks | Aversive odor learning, Figure 5, 6 | Also used for Figure 3 |
| 2016082602 | 14 wks | 23 wks | Aversive odor learning, Figure 5, 6 | - |
| 2017092101 | 15 wks | 26 wks | Neurophysiology recording, Figure 7 |
Additional files
-
Transparent reporting form
- https://doi.org/10.7554/eLife.45658.019
