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

Journal: The Veterinary Quarterly

Article Title: Immunogenicity of replication-deficient vesicular stomatitis virus based rabies vaccine in mice

doi: 10.1080/01652176.2021.1930277

Figure Lengend Snippet: Generation and characterization of VSV/RABV-GP. A: VSV/RABV-GP production was measured by sandwich ELISA. Results have been expressed as the mean of OD450 ± SD values and are representative of at least three independent experiments. Dotted line indicates the background. B: BHK21 cells were transduced with VSV/RABV-GP or VSV/VSV-GP in the presence or absence of the indicated antibodies. Virus entry was quantified by measuring the luciferase levels at 18-h post-transduction. Error bars represent the SD from the mean ( n = 3). Statistical significance was assessed by Student’s t test. * p < 0.05, *** p < 0.001; ns, not significant.

Article Snippet: For sandwich enzyme-linked immunosorbent assay (ELISA), 96-well microplates were coated with mouse monoclonal anti-rabies virus antibody (RV1C5, Santa Cruz) at 1 μg/mL concentration in carbonate/bicarbonate buffer (pH 9.6) and incubated overnight at 4 °C.

Techniques: Sandwich ELISA, Transduction, Virus, Luciferase

Journal: The Veterinary Quarterly

Article Title: Immunogenicity of replication-deficient vesicular stomatitis virus based rabies vaccine in mice

doi: 10.1080/01652176.2021.1930277

Figure Lengend Snippet: Immunization schedule and humoral immune response induced by vaccination. A: Schematic diagram of the immunization protocol. BALB/c mice were i.p. immunized with PBS, RABA, or VSV/RABV-GP at 0 and 14 days after the first immunization. At 28 days post-immunization, mice were sacrificed and serum samples were collected. B and C: Detection of the levels of RABV GP specific IgM (B) and IgG (C). Results are expressed as the mean ( n = 6) of OD450 ± SD values and are representative of at least three independent experiments. Statistical significance was assessed by Student’s t test. *** p < 0.001; ns, not significant.

Article Snippet: For sandwich enzyme-linked immunosorbent assay (ELISA), 96-well microplates were coated with mouse monoclonal anti-rabies virus antibody (RV1C5, Santa Cruz) at 1 μg/mL concentration in carbonate/bicarbonate buffer (pH 9.6) and incubated overnight at 4 °C.

Techniques:

Journal: The Veterinary Quarterly

Article Title: Immunogenicity of replication-deficient vesicular stomatitis virus based rabies vaccine in mice

doi: 10.1080/01652176.2021.1930277

Figure Lengend Snippet: Influence of VSV/RABV-GP immunization on the type of T helper cell response. The serum specific IgG1 (A) and IgG2a (B) titers were determined as indicators of a Th2 or Th2 type of response, respectively. C: IgG1/IgG2a titer ratios of immunized mice. Results are expressed as the mean ( n = 6) of OD450 ± SD values and are representative of at least three independent experiments. Statistical significance was assessed by Student’s t test. ns, not significant.

Article Snippet: For sandwich enzyme-linked immunosorbent assay (ELISA), 96-well microplates were coated with mouse monoclonal anti-rabies virus antibody (RV1C5, Santa Cruz) at 1 μg/mL concentration in carbonate/bicarbonate buffer (pH 9.6) and incubated overnight at 4 °C.

Techniques:

Journal: PLoS ONE

Article Title: Characterization of a Human Antibody Fragment Fab and Its Calcium Phosphate Nanoparticles that Inhibit Rabies Virus Infection with Vaccine

doi: 10.1371/journal.pone.0019848

Figure Lengend Snippet: (A) Proteins immunoprecipitated by Fab094 were separated by SDS-PAGE and probed with mAb(C86307M) by Western blot. (M): protein marker. (1): One protein was recognized by mAb(C86307M). The molecular weight of the protein was about 67 kDa. (2): BHK-21 lysate was used as the negative control to replace Fab094 in the IP. (B): MS spectrums of fragment ions were from the 67 kDa protein. Eight major (m/z = 175.038, 262.012, 362.927, 463.973, 634.074, 737.207, 852.205, 1370.649) ions were detected.

Article Snippet: The commercial mouse anti-rabies virus glycoprotein antibody C86307M, which reacts with a glycoprotein of rabies, more than 20 different strains from 4 serogroups, including CVS, Lagosbat, Mokola, Duwenhage were positive in neutralization (Meridian Life Science, USA), was added to the membranes for 1 h at room temperature.

Techniques: Immunoprecipitation, SDS Page, Western Blot, Marker, Molecular Weight, Negative Control