Cerebellar modulation of memory encoding in the periaqueductal grey and fear behaviour
- School of Physiology, Pharmacology & Neuroscience, University of Bristol, United Kingdom
Figures
Figure 1 with 3 supplements
Single-unit ventrolateral periaqueductal grey (vlPAG) responses during auditory cued fear delay conditioning.
(A) Schematic representation of the fear conditioning paradigm composed of habituation, acquisition, and extinction sessions. Habituation and acquisition were carried out in context A, whilst …
Figure 1—figure supplement 1
Tetrode recordings.
(A) Example electrophysiological data recorded from a ventrolateral periaqueductal grey (vlPAG) tetrode (30 kHz sampling rate, 600–3 kHz bandpass filter, Ch1-4, tetrode channels 1–4). Raw data …
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Figure 1—figure supplement 1—source data 1
Numerical data to support graphs in Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig1-figsupp1-data1-v1.docx
Figure 1—figure supplement 2
Histological verification of implants.
(A) Photomicrographs of two example tetrode tracks (arrows) in periaqueductal grey (PAG). Outlines show left half of two coronal sections (anteroposterior [AP] levels –6.5 and –7.5) of the PAG. …
Figure 1—figure supplement 3
Ventrolateral periaqueductal grey (vlPAG) offset responses during auditory cued fear trace conditioning.
(A) Example type 1 offset unit in vlPAG recorded during extinction following acquisition where a 1 s trace was introduced between conditioned stimulus (CS) offset and the unconditioned stimulus (US) …
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Figure 1—figure supplement 3—source data 1
Numerical data to support graphs in Figure 1—figure supplement 3.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig1-figsupp3-data1-v1.docx
Figure 2 with 1 supplement
Effect of medial cerebellar nucleus (MCN) inactivation during consolidation on ventrolateral periaqueductal grey (vlPAG) type 1 onset and offset responses during extinction.
(A) Schematic timeline representing the fear conditioning protocol for muscimol animals. (B) Group data for control animals showing average z-scored type 1 onset responses (n = 18 single units from …
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Figure 2—source data 1
Numerical data to support graphs in Figure 2.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig2-data1-v1.docx
Figure 2—figure supplement 1
Comparison of single-unit results for tetrodes-only versus saline control animals.
(A) Group data for tetrodes-only control animals (n = 14 units) recorded on day 2, showing average z-scored type 1 onset responses during early extinction (EE, upper panel) and late extinction (LE, …
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Figure 2—figure supplement 1—source data 1
Numerical data to support graphs in Figure 2—figure supplement 1.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig2-figsupp1-data1-v1.docx
Figure 3 with 1 supplement
Auditory event-related field potentials (ERPs) recorded simultaneously in the medial cerebellar nucleus (MCN) and ventrolateral periaqueductal grey (vlPAG) during extinction.
(A) Group average ERPs recorded at CS+ onset in the MCN and vlPAG in control animals (n = 6 rats), arrows indicate time of tone onset; each waveform shows mean ± SEM; dark blue, average ERP during …
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Figure 3—source data 1
Numerical data to support graphs in Figure 3.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig3-data1-v1.docx
Figure 3—figure supplement 1
Relationship between unit activity and event-related potentials (ERPs).
(A) Example ERP recorded in the ventrolateral periaqueductal grey (vlPAG) during early extinction training at CS+ offset. Average of 14 trials. (B) Example single-unit data recorded simultaneously …
Figure 4
Effect of medial cerebellar nucleus (MCN) inactivation during consolidation on the timing of ventrolateral periaqueductal grey (vlPAG) type 1 onset and offset responses during extinction.
(A) Violin plots showing the number of significant peaks of activity (≥2 SD from baseline) that occur in the initial 500 ms following CS+ onset for early extinction (EE) and late extinction (LE) in …
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Figure 4—source data 1
Numerical data to support graphs in Figure 4.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig4-data1-v1.docx
Figure 5 with 2 supplements
Freezing activity during extinction for control and muscimol animals.
(A) The percentage of total time the conditioned tone (CS+) was presented during early extinction (EE) and late extinction (LE) that freezing epochs occurred in control (n = 10 rats) versus muscimol …
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Figure 5—source data 1
Numerical data to support graphs in Figure 5.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig5-data1-v1.docx
Figure 5—figure supplement 1
Trial-by-trial comparison of conditioned behaviour in muscimol and control animals.
(A) Grouped data showing the average percentage of time spent freezing during presentation of each CS+ tone as a function of extinction training. Five blocks of extinction (Ext1–5) with early …
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Figure 5—figure supplement 1—source data 1
Numerical data to support graphs in Figure 5—figure supplement 1.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig5-figsupp1-data1-v1.docx
Figure 5—figure supplement 2
Conditioned fear-related behaviours.
(A) Rate of extinction as measured by the slope of the line of best fit for change in percentage of time spent freezing during presentation of the CS+ over the first 21 presentations of the CS+ …
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Figure 5—figure supplement 2—source data 1
Numerical data to support graphs in Figure 5—figure supplement 2.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig5-figsupp2-data1-v1.docx
Figure 6 with 3 supplements
Effect of medial cerebellar nucleus-periaqueductal grey (MCN-PAG) pathway modulation on behaviour during acquisition.
(A) Schematic timeline representing the fear conditioning protocol for the DREADDs experiment. (B) The effect of clozapine N-oxide (CNO) delivery into the ventrolateral PAG (vlPAG) during …
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Figure 6—source data 1
Numerical data to support graphs in Figure 6.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig6-data1-v1.docx
Figure 6—figure supplement 1
Anatomical mapping of the medial cerebellar nucleus-periaqueductal grey (MCN-PAG) pathway.
(A) Schematic showing a sagittal section of the cerebellum injection into the MCN of an anterograde tracer (AAV-CAG-tdTomato viral vector). To the left, high-power photomicrograph showing …
Figure 6—figure supplement 2
Effect of DREADDs on general motor and affective behaviour.
(A) Histological reconstruction of cannulae placement in periaqueductal grey (PAG) in control (n = 9 rats, tracks colour coded blue to black for different animals) and DREADD animals (n = 10 rats, …
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Figure 6—figure supplement 2—source data 1
Numerical data to support graphs in Figure 6—figure supplement 2.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig6-figsupp2-data1-v1.docx
Figure 6—figure supplement 3
Anatomical evaluation of DREADD transfection of medial cerebellar nucleus-periaqueductal grey (MCN-PAG) pathway.
(A) Schematic showing experimental arrangement for anatomical assessment of DREADD transfection. In each animal, a green fluorescent retrograde tracer (rAAV-CAG-GFP) was injected into the …
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Figure 6—figure supplement 3—source data 1
Numerical data to support graphs in Figure 6—figure supplement 3.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig6-figsupp3-data1-v1.docx
Figure 7 with 1 supplement
Effect of medial cerebellar nucleus-periaqueductal grey (MCN-PAG) pathway modulation on behaviour during extinction.
(A) The percentage of total time the CS+ was presented that animals displayed freezing behaviour during early extinction (EE) (trials 1–14) or late extinction (LE) (trials 21–35) in control (n = 9 …
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Figure 7—source data 1
Numerical data to support graphs in Figure 7.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig7-data1-v1.docx
Figure 7—figure supplement 1
Freezing and ultrasonic vocalisation (USV) behaviour across extinction training.
(A) Grouped data showing the average percentage of time spent freezing during presentation of each CS+ tone as a function of extinction training. Each data point based on average of seven CS+ …
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Figure 7—figure supplement 1—source data 1
Numerical data to support graphs in Figure 7—figure supplement 1.
- https://cdn.elifesciences.org/articles/76278/elife-76278-fig7-figsupp1-data1-v1.docx
Author response image 1
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (species) | Sprague–Dawley (rat) male | Envigo | RRID:RGD_737903 | |
| Transfected construct | pAAV-CAG-tdTomato | Addgene, USA | RRID:Addgene_59462 | Adeno-associated viral vector (AAV1) |
| Transfected construct | rAVV-CAG-GFP | Addgene, USA | RRID:Addgene_37825 | Adeno-associated viral vector (AAVrg) |
| Transfected construct | pAAV-hSyn-hM4D(Gi)-mCherry | Addgene, USA | RRID:Addgene_50475 | Adeno-associated viral vector (AAV5) |
| Transfected construct | pAAV-hSyn-EGFP | Addgene, USA | RRID:Addgene_50465 | Adeno-associated viral vector (AAV5) |
| Chemical compound, drug | Muscimol | Sigma-Aldrich | M1523 | |
| Chemical compound, drug | Clozepine-N-oxide | Tocris Bioscience, UK | 4963 | |
| Antibody | Anti-mCherry (rabbit polyclonal) | BioVision | 5993 | (1:2000) |
| Antibody | Alexa Fluor 594 | Molecular Probes | (1:1000) | |
| Other | CerePlex µ Headstage | Blackrock Microsystems, UT | PN-9716 | |
| Software, algorithm | Blackrock Central Software Suite | Blackrock Microsystems, UT | ||
| Software, algorithm | OBS | Open Broadcaster Software; 2012–2020 | ||
| Software, algorithm | Solomon Coder | András Péter, 2019 | ||
| Software, algorithm | DeepLabCut | Wei and Kording, 2018 | ||
| Software, algorithm | MATLAB | MathWorks | RRID:SCR_001622 | |
| Software, algorithm | Spike7 | Cambridge Electronic Design Limited | ||
| Software, algorithm | NeuroExplorer | Plexon | ||
| Software, algorithm | RStudio | RStudio, USA | ||
| Software, algorithm | Prism 9 | GraphPad, USA |
