Differential inputs to striatal cholinergic and parvalbumin interneurons imply functional distinctions
- The Salk Institute for Biological Studies, United States
Figures
Figure 1 with 2 supplements
Monosynaptic tracing reveals the inputs to striatal ChAT and PV interneurons.
(a) Schematic of Cre-dependent AAV-eGFP viral injection in ChAT-Cre or PV-Cre mice. (b) AAV-eGFP expression is highly colocalized with ChAT and PV immunostaining, respectively. Scale bars, 100 µm. …
Figure 1—figure supplement 1
Overview of inputs to ChAT and PV interneurons.
(a) Schematic of Cre-dependent AAV eGFP injection in striatum of ChAT-Cre or PV-Cre mouse. (b) Quantification of ChAT immunostaining and eGFP expression colocalization. (c) Quantification of PV …
Figure 1—figure supplement 2
ChAT and PV starter cells are restricted to dorsal striatum, similar in total number and are distributed equally across dorsal medial and dorsal lateral striatal subdivisions.
(a) Expression of TVA-mCherry positive starter cells, rabies-eGFP and immunostaining for PV in dorsal striatal injection site. Scale bar, 100 um. (b) Magnified inset of cortex (Ctx) and dorsal …
Figure 2 with 2 supplements
Summary of brain-wide direct inputs to striatal ChAT and PV interneurons.
(a) Major input regions (>0.4% of total inputs) to ChAT (black bar) and PV (red bar) interneurons. A complete set of all inputs to ChAT and PV interneurons is shown in Figure 2—figure supplement 1. …
Figure 2—figure supplement 1
Complete list of all inputs to ChAT and PV interneurons.
(a) All major (>0.4%) and minor (<0.4%) inputs to striatal ChAT (black) and PV (red) interneurons. Short dashed line represents division between major and minor inputs. Mean ± SEM. In blue, *p≤0.05; …
Figure 2—figure supplement 2
Summary of the inputs and proposed function of striatal ChAT and PV interneurons.
Striatal ChAT interneurons receive preferential inputs from orbital cortex, retrosplenial cortex, intralaminar thalamic nuclei, mediodorsal thalamus and the pedunculopontine nucleus and might be …
Figure 3 with 3 supplements
A projection from thalamic reticular nucleus to striatal PV interneurons.
(a) PV interneurons receive a thalamic reticular nucleus (TRN) projection. (b) Rabies-eGFP positive neurons in TRN colocalize with PV immunostaining. Scale bar, 50 µm. Thal = thalamus. (c) …
Figure 3—figure supplement 1
Membrane properties of striatal PV interneurons are similar between TRN connected or non-connected cells.
Rabies positive eGFP cells in TRN colocalize with parvalbumin and somatostatin immunostaining. Somatostatin (Som) positive TRN neurons send projections to striatum. (a–d) Capacitance, membrane …
Figure 3—figure supplement 2
Verification of dorsal striatal in vivo recording array placement.
(a) Cartoon schematic of multichannel array placement with optical fiber for light delivery in dorsal striatum. (b) Grayscale DAPI stained images of striatal slices (left) following in vivo …
Figure 3—figure supplement 3
TRN terminal stimulation inhibits putative striatal fast-spiking (PV) interneurons.
(a) 3D plot of all recorded units by peak half-width (ms), amplitude (mV) and firing rate (Hz). Putative PV interneurons sorted by narrow half widths (<0.12 ms). Red circles denotes putative fast …
Figure 4 with 2 supplements
Functional projections of GPe to striatal ChAT and PV interneurons.
(a) Experimental approach to label GPe inputs with ChR2 and express tdTomato in ChAT or PV interneurons. (b) Averaged paired-pulse (50 ms ISI) light-evoked IPSCs in striatal ChAT interneuron …
Figure 4—figure supplement 1
Cell-type identity of GPe projections to striatal ChAT and PV interneurons.
(a) GPe is the predominant basal ganglia input to ChAT and PV interneurons (two-way ANOVA, F (6,63)=27.27, p<0.0001). (b–c) (top) Few GPe rabies-eGFP neurons colocalize with immunostaining for PV, …
Figure 4—figure supplement 2
GPe functional inhibitory connectivity to ChAT interneurons, PV interneurons and SPNs.
(a) Cartoon schematic showing whole-cell recordings from pairs of interneurons and neighboring SPNs following light-evoked GPe terminal stimulation. (b) IPSCs from pairs of tdTomato (+) ChAT …
Figure 5
Excitatory projections from pedunculopontine nucleus to striatal ChAT interneurons.
(a) Rabies-labeled projections to striatal ChAT interneurons from the PPN are not cholinergic. Note no colocalized eGFP (green) and ChAT immunostaining (red). scp, superior cerebellar peduncle. …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | ChAT-IRES-Cre | Jackson Labs | stock #006410; RRID:IMSR_JAX:006410 | maintained on a C57BL6/J background |
| Strain, strain background (Mus musculus) | PV-Cre | Jackson Labs | stock #008069; RRID:IMSR_JAX:008069 | maintained on a C57BL6/J background |
| Strain, strain background (Mus musculus) | Sst-IRES-Cre | Jackson Labs | stock #028864; RRID:IMSR_JAX:028864 | maintained on a C57BL6/J background |
| Strain, strain background (Adeno-associated virus) | AAV5-TVA-mCherry | UNC Viral Vector Core | RRID: SCR_002448 | 3–4.3 × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAV8-CA-RG | UNC Viral Vector Core | RRID: SCR_002448 | 1.2–4.3 × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | (EnvA) SAD-∆G Rabies-eGFP | Salk Vector Core | RRID: SCR_014847 | 1.6–6.55 × 108 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAV9-Ef1a-DIO-ChR2 (H134R)-eYFP | University of Penn Viral Vector Core | RRID: SCR_015406 | two × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAV9-FLEX-tdTomato | University of Penn Viral Vector Core | RRID: SCR_015406 | two × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAVretro-EF1a-Flp | UNC Viral Vector Core | RRID: SCR_002448 | two × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAV9-hsyn-Con-Fon-hChR2-eYFP | UNC Viral Vector Core | RRID: SCR_002448 | two × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAV9-FLEX-eGFP | University of Penn Viral Vector Core | RRID: SCR_015406 | two × 1012 particles/mL |
| Strain, strain background (Adeno-associated virus) | AAV9-hsyn-ChR2-eYFP | University of Penn Viral Vector Core | RRID: SCR_015406 | two × 1012 particles/mL |
| Antibody | anti-PV (mouse,monoclonal) | MilliporeSigma | P3088; RRID: AB_477329 | 1/1000 |
| Antibody | anti-PV (rabbit, polyclonal) | Abcam | ab11427; RRID: AB_298032 | 1/1000 |
| Antibody | anti-ChAT (goat, polyclonal) | MilliporeSigma | AB144P; RRID: AB_2079751 | 1/100 |
| Antibody | anti-NPAS1 (rabbit, polyclonal) | GeneTex | GTX105876; RRID: AB_424768 | 1/500 |
| Antibody | anti-mCherry (mouse, monoclonal) | Takara Bio USA, Inc (Clontech Labs) | 632543; RRID: AB_2307319 | 1/250 |
| Antibody | anti-eGFP (rabbit, polyclonal) | Thermo Fisher Scientific | A11122; RRID: AB_1074875 | 1/400 |
| Antibody | anti-somatostatin (rabbit, polyclonal) | Protos Biotech Corp | NP106SST | 1/300 |
| Antibody | anti-FoxP2 (rabbit, polyclonal) | Sigma | HPA000382; RRID: AB_1078908 | 1/1000 |
| Antibody | donkey anti-mouse 488 (secondary) | Jackson Immunoresearch | 715-545-150; RRID: AB_2340846 | 1/250 |
| Antibody | donkey anti-mouse CY3 (secondary) | Jackson Immunoresearch | 715-165-150; RRID: AB_2340813 | 1/250 |
| Antibody | donkey anti-mouse CY5 (secondary) | Jackson Immunoresearch | 715-175-150; RRID: AB_2340819 | 1/250 |
| Antibody | donkey anti-rabbit 488 (secondary) | Jackson Immunoresearch | 711-545-152; RRID: AB_2313584 | 1/250 |
| Antibody | donkey anti-rabbit CY3 (secondary) | Jackson Immunoresearch | 711-165-152; RRID: AB_2307443 | 1/250 |
| Antibody | donkey anti-rabbit CY5 (secondary) | Jackson Immunoresearch | 711-175-152; RRID: AB_2340607 | 1/250 |
| Antibody | donkey anti-goat 488 (secondary) | Jackson Immunoresearch | 705-545-147; RRID: AB_2336933 | 1/250 |
| Antibody | donkey anti-goat CY3 (secondary) | Jackson Immunoresearch | 705-165-147; RRID: AB_2307351 | 1/250 |
| Antibody | donkey anti-goat CY5 (secondary) | Jackson Immunoresearch | 705-175-147; RRID: AB_2340415 | 1/250 |
| Chemical compound, drug | NBQX disodium salt hydrate | MilliporeSigma | N183 | 10 uM (final) |
| Chemical compound, drug | DL-APV | MilliporeSigma | A5282 | 50 uM (final) |
| Chemical compound, drug | Picrotoxin | MilliporeSigma | P1675 | 50–100 uM (final) |
| Chemical compound, drug | NMDG | MilliporeSigma | M2004 | |
| Chemical compound, drug | HCl | MilliporeSigma | H1758 | |
| Chemical compound, drug | KCl | MilliporeSigma | P9541 | |
| Chemical compound, drug | NaH2PO4 | MilliporeSigma | S3139 | |
| Chemical compound, drug | NaHCO3 | MilliporeSigma | S6014 | |
| Chemical compound, drug | Glucose | MilliporeSigma | G5767 | |
| Chemical compound, drug | Sodium L-Ascorbate | MilliporeSigma | A4034 | |
| Chemical compound, drug | Sodium Pyruvate | MilliporeSigma | P2256 | |
| Chemical compound, drug | Thiourea | MilliporeSigma | T8656 | |
| Chemical compound, drug | MgSO4 | MilliporeSigma | M2643 | |
| Chemical compound, drug | CaCl2 | MilliporeSigma | 223506 | |
| Chemical compound, drug | MgCl2 | MilliporeSigma | M9272 | |
| Chemical compound, drug | KMeSO4 | MilliporeSigma | 83000 | |
| Chemical compound, drug | HEPES | MilliporeSigma | H4034 | |
| Chemical compound, drug | EGTA | MilliporeSigma | 3777 | |
| Chemical compound, drug | Mg-ATP | MilliporeSigma | A9187 | |
| Chemical compound, drug | Na-GTP | MilliporeSigma | G8877 | |
| Chemical compound, drug | CsCl | MilliporeSigma | C4036 | |
| Chemical compound, drug | CsMeSO3 | MilliporeSigma | C1426 | |
| Chemical compound, drug | QX-314 | MilliporeSigma | L5783 | |
| Chemical compound, drug | TEA-Cl | MilliporeSigma | T2265 | |
| Software, algorithm | MATLAB | RRID: SCR_001622 | ||
| Software, algorithm | GraphPad Prism 6 | RRID: SCR_002798 | ||
| Software, algorithm | Adobe Illustrator CS6 | RRID: SCR_010279 | ||
| Software, algorithm | pClamp9 | RRID: SCR_011323 | ||
| Software, algorithm | Fiji/Imagej | RRID: SCR_002285 | ||
| Other | Allen Reference Atlas | RRID: SCR_013286 |
Additional files
-
Source code 1
MATLAB script for perievent time histogram (PETH) plot.
Plot PETH of ChAT or PV interneuron firing rates aligned to laser on (time zero).
- https://doi.org/10.7554/eLife.35657.016
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Transparent reporting form
- https://doi.org/10.7554/eLife.35657.017
