Journal: eLife

Article Title: An open cortico-basal ganglia loop allows limbic control over motor output via the nigrothalamic pathway

doi: 10.7554/elife.49995

Figure Lengend Snippet: Figure 1. Trans-synaptic wild-type rabies tracing reveals both closed and open cortico-basal ganglia loops. (A) The strategy to label striatal neurons connecting to the cerebral cortex by Wt-RABV trans-synaptic retrograde tracing, and CTb-based non-trans-synaptic anterograde tracing for mapping cortico-striatal terminals. (B) Example image of Wt-RABV/CTb injection into M1 (left). After 66–70 hr of survival time, Wt-RABV was transfected up to 3rd- order neurons, which were found in various striatal subregions (right). Scale bars, 1 mm (left), 500 mm (right). (C) 3D-reconstruction of Wt-RABV+ striatal neurons from the M1 injection case shown in (B). The two different angles emphasize the presence of Wt-RABV+ neurons throughout all of the striatum (VS, DMS, DLS, and TS). (D) Schema of Wt-RABV/CTb injection in M1. (E) Anterogradely labeled CTb+ cortico-striatal terminals (green) and retrogradely labeled Wt-RABV+ striatal neurons (purple) from the M1 injection case shown in (B). (F) Density map showing the distribution of Wt-RABV+ neurons throughout the striatum from M1 injection. Black contours indicate approximate areas receiving cortico-striatal inputs from M1. Color maps indicate the intensity of Wt-RABV+ labeling. (G–I) The same analyses for Wt-RABV/CTb injection in M2. (J–L) The same analyses for Wt-RABV/CTb injection in mPFC. (M) Normalized distribution of Wt-RABV+ neurons across five striatal regions (VMS, VLS, DMS, DLS, TS) showing differences Figure 1 continued on next page

Article Snippet: DOI: https://doi.org/10.7554/eLife.49995 15 of 29 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (recombinant rabies virus) EnvA.dGRabies.eGFP Salk Vector Core RRID: SCR_014847 Strain, strain background (adeno-associated virus) AAV9.FLEX.tdTomato University of Penn Viral Vector Core RRID: SCR_015406 Strain, strain background (adeno-associated virus) AAV9.FLEX.eGFP University of Penn Viral Vector Core RRID: SCR_015406 Strain, strain background (adeno-associated virus) AAV9.CAG.tdTomato UNC Viral Vector Core RRID: SCR_002448 Strain, strain background (adeno-associated virus) AAV9.hsyn.ChR2.eGFP University of Penn Viral Vector Core RRID: SCR_015406 Strain, strain background (adeno-associated virus) AAV5-EF1a-DIOhChR2(H134R)-mCherry University of Penn Viral Vector Core Antibody anti-wild type rabies phosphoprotein mouse monoclonal antibody commercially unavailable (Raux et al., 1997) (1:5000) Antibody anti-cholera toxin b-subunit goat polyclonal antibody List Biological Laboratories Cat.# 704 (1:15000) Antibody anti-tyrosine hydroxylase (TH) mouse monoclonal antibody Millipore Cat.# MAB318 (1:1000) Antibody anti-NeuN rabbit polyclonal antibody Abcam Cat.# ab104225 (1:1000) Antibody anti-GFP chicken polyclonal antibody Novus Biologicals Cat.# NB100-1614 (1:1000) Antibody anti-substance P mouse monoclonal antibody Abcam Cat.# ab14184 (1:1000) Antibody anti-mouse IgG horseradish peroxydase (HRP) (host: rabbit, polyclonal) DAKO Cat.# P260 (1:200) Antibody anti-goat IgG horseradish peroxydase (HRP) (host: rabbit, polyclonal) DAKO Cat.# P044901-2 (1:200) Antibody anti-mouse Alexa Fluor 488 (host: donkey, polyclonal) Jackson ImmunoResearch Laboratories Cat.# 715-545-151 (1:250) Antibody anti-mouse Cy3 (host: donkey, polyclonal) Jackson ImmunoReseach Laboratories Cat.# 715-165-151 (1:250) Antibody anti-mouse Cy5 (host: donkey, polyclonal) Jackson ImmunoReseach Laboratories Cat.# 715-175-151 (1:250) Continued on next page Aoki et al. eLife 2019;8:e49995.

Techniques: Retrograde Tracing, Anterograde Tracing, Injection, Transfection, Labeling