Journal: Science Advances
Article Title: Neuromodulatory control of inhibitory network arborization in the developing postnatal neocortex
Figure Lengend Snippet: ChC axonal arborization requires Cav3.3-T-type VDCC genes in a dosage sensitive manner. ( A and B ) Confocal images of EP13 transplanted ZsGreen-labeled ChCs transfected with LacZ sgRNAs (A, left) or Cav3.3 sgRNAs (A, right) as well as EP13 transplanted GFP-labeled ChCs originating from Cav3.3 germline KO mice (B). Single optical sections indicating synaptic cartridges apposed to AnkG-labeled AISs are shown in the middle panels. The areas in white boxes with arrowheads are enlarged and overlaid with the AnkG channel. Bottom panels indicate reconstructed axonal arbors in the area defined in Fig. 4A . ( C ) The number of branch points in LacZ sgRNA –transfected ChCs ( n = 5 cells from three mice), Cav3.3 sgRNA– transfected homozygous KO ChCs ( n = 5 cells from three mice), transplanted ChCs originating from Cav3.3 homozygous KO mice ( n = 6 cells from three mice), and transplanted ChCs originating from Cav3.3 heterozygous KO mice ( n = 5 cells from three mice). One-way ANOVA, * P
Article Snippet: The following primary antibodies were used: goat anti-ChAT (1:200; Millipore, catalog no. AB144P), rabbit anti-Cav3.1 (1:100; Alomone Labs, catalog no.ACC-021), rabbit anti-Cav3.2 (1:100; Alomone Labs, catalog no. ACC-025), rabbit anti-Cav3.3 (1:100; Alomone Labs, catalog no. ACC-009), guinea pig polyclonal anti-PV (1:2000; Swant, PVG-213), rat anti-hemagglutinin (1:500; Roche, catalog no.11-867-423-001), chicken anti-GFP (1:800; Abcam, catalog no.ab13970), mouse anti-AnkG (1:500; UC Davis/NIH Neuromab, catalog no. clone N106/36 75-146), rabbit anti-ZsGreen (1:500; Clontech, catalog no. 632474), and rabbit anti-RFP (1:800; Rockland, catalog no. 600-401-379).
Techniques: Labeling, Transfection, Mouse Assay