Differential inputs to striatal cholinergic and parvalbumin interneurons imply functional distinctions

5 figures, 1 table and 2 additional files

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. …

https://doi.org/10.7554/eLife.35657.002
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 …

https://doi.org/10.7554/eLife.35657.003
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 …

https://doi.org/10.7554/eLife.35657.004
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. …

https://doi.org/10.7554/eLife.35657.005
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; …

https://doi.org/10.7554/eLife.35657.006
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 …

https://doi.org/10.7554/eLife.35657.007
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) …

https://doi.org/10.7554/eLife.35657.008
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 …

https://doi.org/10.7554/eLife.35657.009
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

https://doi.org/10.7554/eLife.35657.010
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 …

https://doi.org/10.7554/eLife.35657.011
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 …

https://doi.org/10.7554/eLife.35657.012
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, …

https://doi.org/10.7554/eLife.35657.013
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 …

https://doi.org/10.7554/eLife.35657.014
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. …

https://doi.org/10.7554/eLife.35657.015

Tables

Key resources table
Reagent type (species)
or resource
DesignationSource or referenceIdentifiersAdditional
information
Strain, strain background (Mus musculus)ChAT-IRES-CreJackson Labsstock #006410; RRID:IMSR_JAX:006410 maintained on a C57BL6/J background
Strain, strain background (Mus musculus)PV-CreJackson Labsstock #008069; RRID:IMSR_JAX:008069maintained on a C57BL6/J background
Strain, strain background (Mus musculus)Sst-IRES-CreJackson Labsstock #028864; RRID:IMSR_JAX:028864maintained on a C57BL6/J background
Strain, strain background (Adeno-associated virus)AAV5-TVA-mCherryUNC Viral Vector CoreRRID: SCR_0024483–4.3 × 1012 particles/mL
Strain, strain background (Adeno-associated virus)AAV8-CA-RGUNC Viral Vector CoreRRID: SCR_0024481.2–4.3 × 1012 particles/mL
Strain, strain background (Adeno-associated virus)(EnvA) SAD-∆G Rabies-eGFPSalk Vector CoreRRID: SCR_0148471.6–6.55 × 108 particles/mL
Strain, strain background (Adeno-associated virus)AAV9-Ef1a-DIO-ChR2 (H134R)-eYFPUniversity of Penn Viral Vector CoreRRID: SCR_015406two × 1012 particles/mL
Strain, strain background (Adeno-associated virus)AAV9-FLEX-tdTomatoUniversity of Penn Viral Vector CoreRRID: SCR_015406two × 1012 particles/mL
Strain, strain background (Adeno-associated virus)AAVretro-EF1a-FlpUNC Viral Vector CoreRRID: SCR_002448two × 1012 particles/mL
Strain, strain background (Adeno-associated virus)AAV9-hsyn-Con-Fon-hChR2-eYFPUNC Viral Vector CoreRRID: SCR_002448two × 1012 particles/mL
Strain, strain background (Adeno-associated virus)AAV9-FLEX-eGFPUniversity of Penn Viral Vector CoreRRID: SCR_015406two × 1012 particles/mL
Strain, strain background (Adeno-associated virus)AAV9-hsyn-ChR2-eYFPUniversity of Penn Viral Vector CoreRRID: SCR_015406two × 1012 particles/mL
Antibodyanti-PV (mouse,monoclonal)MilliporeSigmaP3088; RRID: AB_4773291/1000
Antibodyanti-PV (rabbit, polyclonal)Abcamab11427; RRID: AB_2980321/1000
Antibodyanti-ChAT (goat, polyclonal)MilliporeSigmaAB144P; RRID: AB_20797511/100
Antibodyanti-NPAS1 (rabbit, polyclonal)GeneTexGTX105876; RRID: AB_4247681/500
Antibodyanti-mCherry (mouse, monoclonal)Takara Bio USA, Inc (Clontech Labs)632543; RRID: AB_23073191/250
Antibodyanti-eGFP (rabbit, polyclonal)Thermo Fisher ScientificA11122; RRID: AB_10748751/400
Antibodyanti-somatostatin (rabbit, polyclonal)Protos Biotech CorpNP106SST1/300
Antibodyanti-FoxP2 (rabbit, polyclonal)SigmaHPA000382; RRID: AB_10789081/1000
Antibodydonkey anti-mouse 488 (secondary)Jackson Immunoresearch715-545-150; RRID: AB_23408461/250
Antibodydonkey anti-mouse CY3 (secondary)Jackson Immunoresearch715-165-150; RRID: AB_23408131/250
Antibodydonkey anti-mouse CY5 (secondary)Jackson Immunoresearch715-175-150; RRID: AB_23408191/250
Antibodydonkey anti-rabbit 488 (secondary)Jackson Immunoresearch711-545-152; RRID: AB_23135841/250
Antibodydonkey anti-rabbit CY3 (secondary)Jackson Immunoresearch711-165-152; RRID: AB_23074431/250
Antibodydonkey anti-rabbit CY5 (secondary)Jackson Immunoresearch711-175-152; RRID: AB_23406071/250
Antibodydonkey anti-goat 488 (secondary)Jackson Immunoresearch705-545-147; RRID: AB_23369331/250
Antibodydonkey anti-goat CY3 (secondary)Jackson Immunoresearch705-165-147; RRID: AB_23073511/250
Antibodydonkey anti-goat CY5 (secondary)Jackson Immunoresearch705-175-147; RRID: AB_23404151/250
Chemical compound, drugNBQX disodium salt hydrateMilliporeSigmaN18310 uM (final)
Chemical compound, drugDL-APVMilliporeSigmaA528250 uM (final)
Chemical compound, drugPicrotoxinMilliporeSigmaP167550–100 uM (final)
Chemical compound, drugNMDGMilliporeSigmaM2004
Chemical compound, drugHClMilliporeSigmaH1758
Chemical compound, drugKClMilliporeSigmaP9541
Chemical compound, drugNaH2PO4MilliporeSigmaS3139
Chemical compound, drugNaHCO3MilliporeSigmaS6014
Chemical compound, drugGlucoseMilliporeSigmaG5767
Chemical compound, drugSodium L-AscorbateMilliporeSigmaA4034
Chemical compound, drugSodium PyruvateMilliporeSigmaP2256
Chemical compound, drugThioureaMilliporeSigmaT8656
Chemical compound, drugMgSO4MilliporeSigmaM2643
Chemical compound, drugCaCl2MilliporeSigma223506
Chemical compound, drugMgCl2MilliporeSigmaM9272
Chemical compound, drugKMeSO4MilliporeSigma83000
Chemical compound, drugHEPESMilliporeSigmaH4034
Chemical compound, drugEGTAMilliporeSigma3777
Chemical compound, drugMg-ATPMilliporeSigmaA9187
Chemical compound, drugNa-GTPMilliporeSigmaG8877
Chemical compound, drugCsClMilliporeSigmaC4036
Chemical compound, drugCsMeSO3MilliporeSigmaC1426
Chemical compound, drugQX-314MilliporeSigmaL5783
Chemical compound, drugTEA-ClMilliporeSigmaT2265
Software, algorithmMATLABRRID: SCR_001622
Software, algorithmGraphPad Prism 6RRID: SCR_002798
Software, algorithmAdobe Illustrator CS6RRID: SCR_010279
Software, algorithmpClamp9RRID: SCR_011323
Software, algorithmFiji/ImagejRRID: SCR_002285
OtherAllen Reference AtlasRRID: 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
Transparent reporting form
https://doi.org/10.7554/eLife.35657.017

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