Purkinje cell outputs selectively inhibit a subset of unipolar brush cells in the input layer of the cerebellar cortex
- Department of Neurobiology, Harvard Medical School, United States
Figures
Figure 1 with 1 supplement
PC collateral synapses colocalized with mGluR1 UBCs in the vestibular lobules.
(A) Maximal intensity projection of a 15-µm confocal z-stack images of PC collateral synapses co-labeled by synaptophysin-tdTomato (red) and VGAT (green). (B) Annotated PC collateral synapses (cyan) …
Figure 1—figure supplement 1
Automatic synapse detection using a convolutional neural network.
(A) Raw image of PC collateral synapses co-labeled by synaptophysin-tdTomato (red) and VGAT (green). mGluR1 labeling (not shown) was also used as input to the network as it provided additional …
Figure 2 with 1 supplement
PC collaterals preferentially inhibit mGluR1+ UBCs via fast GABAAR-mediated feedback.
(A) Schematic showing a mossy fiber (MF) and a PC collateral innervating a UBC. A whole-cell electrode was used to voltage clamp the UBC, and responses were measure for either a glutamate puff …
Figure 2—figure supplement 1
GABAA and GABAB receptor subunits are preferentially expressed in mGluR1-positive UBCs.
Molecular data is from https://singlecell.broadinstitute.org/single_cell/study/SCP795/. We analyzed scRNAseq data of UBCs for GABAA, glycine, and GABAB receptor subunits expression levels in …
Figure 3
Paired recordings of PC to UBC synaptic connections.
(A) Schematic of the paired recording configuration. (B) A representative pair showing a cell-attached PC recording (top) and whole-cell voltage-clamp recording of a synaptically connected UBC …
Figure 4 with 1 supplement
PC collaterals provide slow GABAB receptor-mediated inhibition onto lobule X UBCs.
(A) Schematic of full-field optical stimulation for modulating PC firing rate in lobule X. (B) Cell-attached recording of spontaneous PC spikes for no (top), low intensity (middle), and high …
Figure 4—figure supplement 1
Perforated patch recordings provide stable responses to a GABAB-receptor agonist.
(A) Sample whole-cell (top, gray) and perforated (bottom, black) responses to baclofen puffs immediately after break-in (left) and 8 min later (right column). (B) Baclofen response amplitudes …
Figure 5
PC-UBC feedback via GABABR sharpens temporal response to current injections.
(A) Sample perforated current-clamp recording of UBC spiking response to an 8 pA current step (500 ms), without (gray, left) and with a hyperpolarizing tonic current (black, right). (B) Average …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | C57BL/6 | Charles River | ||
| Strain, strain background (M. musculus) | B6.Cg-Tg(Pcp2-cre)3555Jdhu/J | Jackson Laboratory | Strain#010536 | |
| Strain, strain background (M. musculus) | ChR2-EYFP (Ai32) | Jackson Laboratory | Strain#024109 | |
| Strain, strain background (M. musculus) | Synaptophysin-tdTomato (Ai34D) | Jackon Laboratory | Strain#021570 | |
| Chemical compound, drug | NBQX disodium salt | Abcam | Ab120046 | |
| Chemical compound, drug | (R)-CPP | Abcam | Ab120159 | |
| Chemical compound, drug | Strychnine hydrochloride | Abcam | Ab120416 | |
| Chemical compound, drug | SR95531 (Gabazine) | Abcam | Ab120042 | |
| Chemical compound, drug | L-Glutamate | Abcam | Ab120049 | |
| Chemical compound, drug | (RS)-3,5-DHPG | Tocris | Cat. no. 0342/1 | |
| Chemical compound, drug | (R,S)-Baclofen | Abcam | Ab120149 | |
| Chemical compound, drug | CGP 55845 hydrochloride | Tocris | Cat. no. 1248 | |
| Antibodies | Guinea pig anti-VGAT (Guinea pig polyclonal) | Synaptic Systems | 131004 | 1 μg/mL stock, 1:200 |
| Antibodies | Mouse anti-rat mGluR1 (Mouse monoclonal) | BD Pharmingen | Cat. no. 556331 | 0.5 mg/mL stock, 1:800 |
| Antibodies | Goat anti-Guinea pig Alexa Fluor 488 (Goat polyclonal) | Abcam | Ab150185 | 1:500 |
| Antibodies | Goat anti-mouse Alexa Fluor 647 (Goat poly clonal) | Invitrogen | A32728 | 1:500 |
| Software, algorithm | Igor Pro 6 | Wavemetrics | https://www.wavemetrics.com/ | |
| Software, algorithm | MafPC | Courtesy of M.A. Xu-Friedman | https://www.xufriedman.org/mafpc | |
| Software, algorithm | MATLAB (R2017a) | MathWorks | https://www.mathworks.com/products/matlab.html | |
| Software, algorithm | ImageJ | NIH | https://imagej.nih.gov/ij/ | |
| Software, algorithm | Python | Python Software Foundation | https://www.python.org/ | |
| Software, algorithm | Pytorch | Opensource | https://www.pytorch.org/ |
