Parabrachial CGRP neurons modulate active defensive behavior under a naturalistic threat
- School of Psychology, Korea University, Republic of Korea
Figures
Figure 1 with 1 supplement
Distinct firing response patterns of calcitonin gene-related peptide (CGRP) neurons to different aversive stimuli.
(A) Schematic of AAV-DIO-ChR2-mScarlet injection and optrode implantation into the parabrachial nucleus (PBN) of CalcaCre/+ mice (n=6) and the corresponding representative histology image. (B) …
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Figure 1—source data 1
Behavior data for Figure 1.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig1-data1-v2.xlsx
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Figure 1—source data 2
Electrophysiology data for Figure 1.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig1-data2-v2.xlsx
Figure 1—figure supplement 1
Characterization of CGRP neuron activity and behavioral responses during fear conditioning with chasing robot.
(A) Reconstruction of optrode implantation sites along the anterior-posterior extent of the parabrachial nucleus (PBN).Each bar indicates an optrode track included for data analysis in the …
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Figure 1—figure supplement 1—source data 1
Correlations between spontaneous and light-evoked waveforms.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig1-figsupp1-data1-v2.xlsx
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Figure 1—figure supplement 1—source data 2
Velocity data for a representative animal across 10 fear conditioning trials.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig1-figsupp1-data2-v2.xlsx
Figure 2
Stimulation of calcitonin gene-related peptide (CGRP) neurons in the absence of any external stimuli induces robust freezing behavior.
(A) Schematic of bilateral AAV-DIO-ChR2-mScarlet or AAV-DIO-Jaws-GFP injections and optic fiber implantation into the parabrachial nucleus (PBN), with representative histological images of viral …
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Figure 2—source data 1
Behavior data for Figure 2.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig2-data1-v2.xlsx
Figure 3 with 1 supplement
Activation of calcitonin gene-related peptide (CGRP) neurons during unconditioned stimulus (US) presentation enhances active defensive behavior.
(A) A schematic diagram of fear conditioning protocol with the robot. CGRP neuronal activity was bidirectionally manipulated during the presentation of the robot chasing. (B) Freezing to the …
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Figure 3—source data 1
Behavior data for Figure 3.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Optic fiber placements and behavioral assessments during conditioning and retention test.
(A–B) Anatomical locations of optic fiber tips above the parabrachial nucleus (PBN) of (A) ChR2 and (B) Jaws groups.(C) Number of times bumping with the robot. No group differences were observed …
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Figure 3—figure supplement 1—source data 1
Number of bumping incidents across different groups and velocity data for Figure 3—figure supplement 1D.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig3-figsupp1-data1-v2.xlsx
Figure 4
Calcitonin gene-related peptide (CGRP) activation during conventional fear conditioning promotes passive, but not active, defensive behavior.
(A) A schematic diagram of fear conditioning protocol with the footshock. (B) Freezing to the conditioned stimulus (CS) during habituation and conditioning sessions. All three groups showed a …
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Figure 4—source data 1
Behavior data for Figure 4.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig4-data1-v2.xlsx
Figure 5 with 1 supplement
Escalating threat intensity modulates defensive behavior.
(A) Freezing to the conditioned stimulus (CS) during habituation and conditioning sessions for groups subjected to three different robot speeds (n=8 per group). All three groups showed an equivalent …
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Figure 5—source data 1
Behavior data for Figure 5.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Effects of robot speed on bumping incidents and behavioral responses.
(A) Number of times bumping with the robot at different speeds.Bumping occurrence increased with robot speeds (significant group effect in a mixed-design ANOVA, F(2, 21)=29.44, p<0.001): 80 cm/s …
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Figure 5—figure supplement 1—source data 1
Number of bumping incidents across groups at different robot speeds and velocity data for Figure 5—figure supplement 1C.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig5-figsupp1-data1-v2.xlsx
Figure 6 with 1 supplement
Calcitonin gene-related peptide (CGRP) neurons are necessary for promoting active defensive behaviors under high-speed threat conditions.
(A) Schematic of bilateral AAV-DIO-GFP:TetTox or AAV-DIO-eYFP (control) injections into the parabrachial nucleus (PBN) and representative histological images of TetTox expression. (B) Freezing to …
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Figure 6—source data 1
Behavior data for Figure 6.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Effects of CGRP neuron silencing on bumping and CS-evoked velocity.
(A) Number of times bumping with the robot.No group differences were observed throughout the trials (repeated-measures two-way ANOVA, F(1, 18)=0.41, p=0.84). (B) Average velocities in response to …
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Figure 6—figure supplement 1—source data 1
Number of bumping incidents and velocity data for Figure 6—figure supplement 1B.
- https://cdn.elifesciences.org/articles/101523/elife-101523-fig6-figsupp1-data1-v2.xlsx
Author response image 1
Videos
Video 1
Fleeing behavior during conditioning in the ChR2 group in response to conditioned stimulus (CS).
The video shows the behavior of a representative animal from the control, ChR2, and Jaws groups during the last trial of conditioning. The sequence includes 10 s pre-CS, 10 s of CS (7 s of CS alone …
Video 2
Fleeing behavior during aversive memory recall in the ChR2 group in response to conditioned stimulus (CS).
To test the fear memory in the retention test, we used a rectangular box instead of the donut maze used during conditioning. The box was placed on top of the donut maze, preventing the animals from …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | B6.Cg-Calcatm1.1(cre/EGFP)Rpa/J | The Jackson Laboratory | RRID:IMSR_JAX:033168 | |
| Strain, strain background (C57BL/6 J, C57BL/6 J) | C57BL/6 J | The Jackson Laboratory | RRID:IMSR_JAX:000664 | |
| Recombinant DNA reagent | AAV-DIO-ChR2-mScarlet | IBS Virus Facility | N/A | |
| Recombinant DNA reagent | AAV-DIO-Jaws-GFP | Jo et al., 2018 | N/A | |
| Recombinant DNA reagent | AAV-DIO-eYFP | Addgene | RRID:Addgene_27056 | |
| Recombinant DNA reagent | AAV-DIO-TetTox-GFP | Han et al., 2015 | N/A | |
| Software, algorithm | ANY-maze 5.3 | Stoelting Co | N/A | |
| Software, algorithm | GraphPad Prism | GraphPad software | N/A | |
| Software, algorithm | SPSS | IBM | N/A | |
| Software, algorithm | Offline Sorter | Plexon Inc | N/A |
Additional files
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MDAR checklist
- https://cdn.elifesciences.org/articles/101523/elife-101523-mdarchecklist1-v2.docx
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Supplementary file 1
Statistical summary of behavioral and neural data.
- https://cdn.elifesciences.org/articles/101523/elife-101523-supp1-v2.xlsx
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Source code 1
Arduino code for controlling the chasing robot.
- https://cdn.elifesciences.org/articles/101523/elife-101523-code1-v2.zip
