Monosynaptic tracing maps brain-wide afferent oligodendrocyte precursor cell connectivity
- Stanford University, United States
Figures
Figure 1 with 1 supplement
Monosynaptically-restricted rabies virus enables tracing of synaptic inputs to OPCs.
(a) Outline of experimental strategy used to label inputs to Pdgfra+ OPCs. (b) Injection of SADΔG-EGFP(EnvA) into sub-cingular corpus callosum results in widespread labeling of EGFP+ input neurons …
Figure 1—figure supplement 1
SADΔG-EGFP(EnvA) specificity and starter characterization.
(a) Immunofluorescence staining for the white matter and reactive astrocyte marker Gfap (white; top panels) and the macrophage marker Iba1 (white; bottom panels) does not identify substantial …
Figure 2 with 1 supplement
Neuronal inputs to callosal OPCs arise from functionally interconnected cortical and thalamic areas.
(a) Representative sections of neuronal input labeling to OPCs following stereotaxic injection of SADΔG-EGFP(EnvA) to corpus callosum underlying the secondary motor area. Green = EGFP, white = DAPI. …
Figure 2—figure supplement 1
PSD95 puncta colocalization with OPCs.
(a) Representative micrograph of PSD95 puncta (red) and Pdgfra+ (green) OPC in corpus callosum, with subsequent rendering of 3D filament model of OPC and colocalized puncta generated in Imaris. …
Figure 3
Circuit-specific cortical and thalamic neuronal inputs to OPCs in secondary motor area (MOs).
(a) Representative sections of neuronal input labeling to OPCs following stereotaxic injection of SADΔG-EGFP(EnvA) to MOs. Green = EGFP, white = DAPI. (b) Schematic of injection site. (c) Linear …
Figure 4
Circuit-specific cortical and thalamic neuronal neuronal inputs to SSp.
(a) Representative sections of neuronal input labeling to OPCs following stereotaxic injection of SADΔG-EGFP(EnvA) to SSp. Green = EGFP, white = DAPI. (b) Schematic of injection site. (c) Linear …
Figure 5
Thalamic inputs to OPCs arise from functionally-related nuclei.
Tiled immunofluorescence images of GFP+ input neurons were registered to the Allen Brain Atlas to determine the thalamic nuclei from which the inputs arise. Nuclei are color-coded here according to …
Figure 6
Neuronal input to OPC starter ratios are consistent across brain region and despite whisker trimming-induced afferent activity deprivation.
(a) Neuronal input to OPC starter ratios, as measured by the slope of the best-fit linear regression of GFP+ input neurons against Pdgfra+/GFP+ OPC starter cells. Bars indicate mean, error bars …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Anti-Pdgfra (goat polyclonal) | R and D systems | AF1062 RRID:AB_2236897 | 1:500 |
| Antibody | Anti-Olig2 (rabbit monoclonal) | Abcam | EPR2673 RRID:AB_10861310 | 1:500 |
| Antibody | Anti-GFP (chicken polyclonal) | Abcam | Ab13970 RRID:AB_300798 | 1:1000 |
| Antibody | Anti-parvalbumin (rabbit polyclonal) | Abcam | Ab11427 RRID:AB_298032 | 1:250 |
| Antibody | Anti-somatostatin (rat monoclonal) | Millipore | MAB354 RRID:AB_2255365 | 1:200 |
| Antibody | Anti-VIP (rabbit polyclonal) | Immunostar | 20077 RRID:AB_572270 | 1:500 |
| Antibody | Anti-Iba1 (rabbit polyclonal) | Wako | 019–19741 RRID:AB_839504 | 1:500 |
| Antibody | Anti-Cre recombinase (mouse monoclonal) | Millipore | MAB3120 RRID:AB_2085748 | 1:1000 |
| Antibody | Anti-PSD95 (rabbit polyclonal) | Invitrogen | 51–6900 RRID:AB_2533914 | 1:100 |
| Antibody | Anti-CASPR (rabbit monoclonal) | Cell Signaling Technologies | 97736 RRID:AB_2800288 | 1:250 |
| Software | Imaris | Bitplane/Oxford Instruments | v8.1.2 RRID:SCR_007370 | |
| Software | Matlab | Mathworks | R2017b RRID:SCR_001622 | |
| Strain (Mus musculus) | B6N.Cg-Tg(Pdgfra-cre/ERT)467Dbe/J | The Jackson Laboratory | 018280 RRID:IMSR_JAX:018280 | |
| Strain (Mus musculus) | B6;129P2-Gt(ROSA)26Sortm1(CAG-RABVgp4,-TVA)Arenk/J | The Jackson Laboratory | 024708 RRID:IMSR_JAX:024708 |
Additional files
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Source data 1
Raw data presented in the paper.
- https://doi.org/10.7554/eLife.49291.010
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Transparent reporting form
- https://doi.org/10.7554/eLife.49291.011
