plasmid for envacoated cvs-strain glycoprotein-deleted rabies virus fused with egfp (envacvs-n2cδg-egfp)  (Addgene inc)

Journal: bioRxiv

Article Title: A Novel Single Vector Intersectional AAV Strategy for Interrogating Cellular Diversity and Brain Function

doi: 10.1101/2023.02.07.527312

Figure Lengend Snippet: a , ConVERGD-based rabies helper AAVs expressing TVA receptor (magenta) and N2c glycoprotein (brown) were unilaterally co-injected into the LC of Pdyn Cre ; Dbh Flp mice three weeks prior to injection of RABV-CVS-N2c ΔG -H2B-eGFP (green). b , Representative images of TVA(mCh) expression (magenta) and RABV-GFP expression (green) within the LC (TH, white). TVA(mCh) is present only in a subset of LC cells. The inset image highlights a TH + LC starter cell (yellow arrow) containing GFP expression from the rabies virus and mCherry expression from ConVERGD-TVA(mCh). c , Quantitative distribution of GFP + cells across the brain. Highlighted regions represent areas containing at least 3% of total GFP + cells counted. Colors correspond to the regions specified in d ,. The grey cross-hatched region represents the area around the LC that was removed from quantification. Light gray traces represent the distribution of GFP + cell labeling from individual brains. The bold line represents an average of all brains. n = 5 mice. d, Representative images of GFP + cells in major input regions. Cell nuclei are counterstained with DAPI (blue). e, Representative images of axon projections from TVA(mCh) + (magenta) LC cells. Nuclei are counterstained with DAPI (blue). PGRNl/d, paragigantocellular reticular nucleus lateral/dorsal region; PAG, periaqueductal gray; MRN, midbrain reticular nucleus; CeA, central amygdala; LHA, lateral hypothalamic area; BNST, bed nucleus of the stria terminalis; MPO, medial preoptic area of the hypothalamus; PLC, prelimbic cortical area; OFC, orbitofrontal cortex; HC, hippocampus; OB, olfactory bulb. Scale bars are 100μm in b, d, and e. Inset scale bar in b, is 25μm. TH - tyrosine hydroxylase; mChr - mCherry; LC - locus coeruleus; Pdyn - prodynorphin; Dbh - dopamine-β-hydroxylase.

Article Snippet: RABV-CVS-N2c ΔG -H2B-eGFP rabies virus was propagated in the Schwarz lab from an aliquot obtained from the Allen Institute for Brain Science using cell lines and protocols described in Reardon et al.

Techniques: Expressing, Injection, Virus, Labeling