Data from: Characterization of direct Purkinje cell outputs to the brainstem
Data files
Feb 19, 2025 version files 61.86 GB
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code_dir.zip
476.27 MB
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figure1_slidescanner.zip
82.39 MB
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README.md
2.86 KB
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sld1slc12_multiCh.tif
5.17 GB
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Sld1slc12-left-TileScan_1_Region_2_Merged_z00_ch03.tif
1.19 GB
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sld2slc1__multiCh.tif
6.90 GB
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Sld2slc1-TileScan_1_Region_1_Merged_z00_ch03.tif
3.45 GB
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sld2slc8__multiCh.tif
9.38 GB
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Sld2slc8-TileScan_6_Region_1_Merged_z00_ch03.tif
4.69 GB
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sld3slc11_multiCH_ch04.tif
5.73 GB
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Sld3Slc11-TileScan_1_Region_5_Merged_z00_ch03.tif
5.73 GB
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Sld3Slc2-Slide3_101722_Region_3_Merged_z00_ch03.tif
4.75 GB
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sld3slc8__multiCh.tif
5.81 GB
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Sld3Slc8-Region_2_Merged_RAW_ch03.tif
2.90 GB
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sld3sld2_multiCh_ch04.tif
5.59 GB
Abstract
Purkinje cells (PCs) primarily project to cerebellar nuclei but also directly innervate the brainstem. Some PC-brainstem projections have been described previously, but most have not been thoroughly characterized. Here we use a PC-specific cre line to anatomically and electrophysiologically characterize PC projections to the brainstem. PC synapses are surprisingly widespread, with the highest densities found in the vestibular and parabrachial nuclei. However, there are pronounced regional differences in synaptic densities within both the vestibular and parabrachial nuclei. Large optogenetically-evoked PC-IPSCs are preferentially observed in subregions with the highest densities of PC synapses, suggesting that PCs selectively influence these areas and the behaviors they regulate. Unexpectedly, the pontine central gray and nearby subnuclei also contained a low density of PC synapses, and large PC-IPSCs are observed in a small fraction of cells. We combined electrophysiological recordings with immunohistochemistry to assess the molecular identities of these PC targets. PC synapses onto mesencephalic trigeminal neurons were not observed even though these cells are in close proximity to PC boutons. PC synapses onto locus coeruleus neurons are exceedingly rare or absent, even though previous studies concluded that PCs are a major input to these neurons. The availability of a highly selective cre line for PCs allowed us to study functional synapses, while avoiding complications that can accompany the use of viral approaches. We conclude that PCs directly innervate numerous brainstem nuclei, but only inhibit a small fraction of cells in many nuclei. This suggests that PCs target cell types with specific behavioral roles in brainstem regions.
https://doi.org/10.5061/dryad.s7h44j1hw
Description of the data and file structure
This data set are example images and analysis used in the published work, "Characterization of direct Purkinje cell outputs to the brainstem eLife 13:RP101825"
https://doi.org/10.7554/eLife.101825.1
Files and variables
File: figure1_slidescanner.zip
Description:
- raw images used for figure 1
- sorted by file name, coronal slides are named posterior to anterior, sagittal slices are named lateral to medial
File: code_dir.zip
Description:
- analysis scripts for figure 2-4
- excel tables are exports from arivis vgat putative synapse location
- put raw confocal images (see below) into the same directory as the rest of code_dir to run ‘fig2_colocalization.m’ - this script takes vat location data and synaptophysin confocal data and calculate colocalization
- 'fig2_colocalization.m’ is a matlab script takes vat location data in excel and synaptophysin confocal data in raw tiff files and calculate colocalization
- ’fig3_ephys_plot.m’ is a matlab script that takes variable from ‘figure3_ephys_varaible.mat’ and plot cell response amplitude on allen atlas outlines.
- ‘figure3_ephys_varaible.mat’ is a matlab variable file that include two structure variable, each corresponding to cell location and cell amplitude.
- ’fig4_ephys_plot.m’ is a matlab script that takes variable from ‘figure4_ephys_varaible_LC.mat’ and plot cell response amplitude on allen atlas outlines.
- ‘figure4_ephys_varaible_LC.mat’ is a matlab variable file that include two structure variable, each corresponding to cell location and cell amplitude.
- ‘masterslice’ is a directory that contain jpg files of allen atlas outlines of the dorsal brainstem at bregma -5.65mm, -6.05mm, -6.35mm, -6.65mm
File: Sld1slc12-left-TileScan_1_Region_2_Merged_z00_ch03.tif
File: Sld2slc1-TileScan_1_Region_1_Merged_z00_ch03.tif
File: Sld3Slc8-Region_2_Merged_RAW_ch03.tif
File: Sld2slc8-TileScan_6_Region_1_Merged_z00_ch03.tif
File: Sld3Slc2-Slide3_101722_Region_3_Merged_z00_ch03.tif
File: Sld3Slc11-TileScan_1_Region_5_Merged_z00_ch03.tif
- The above list of raw syanptophysin channel confocal image stack used for figure 2 colocalization analysis
File: sld1slc12_multiCh.tif
File: sld3slc8__multiCh.tif
File: sld2slc1__multiCh.tif
File: sld2slc8__multiCh.tif
File: sld3slc11_multiCH_ch04.tif
File: sld3sld2_multiCh_ch04.tif
list of raw synaptophysin channel and vat channel confocal image that generated vGAT location using ZEISS arivis
