Figure 4.

Striatal injection of AAV-Cre leads to a specific decrease of beta2 mRNA in the striatum of beta2-flox/flox mice. a, Top, Diagram showing exon 5 of the Chrnb2 gene flanked by loxP sites in beta2-flox/flox mice. Locations of the three primers used in the ELFO analysis below are also indicated. Bottom, The ELFO analysis shows samples from two representative mice injected with AAV-GFP or AAV-Cre-GFP in the DS. Bottom row on the gel shows 200 bp products of #198 and #199 primers. Top additional band shows an additional 400 bp product of #198 and Bot2-09 primers indicating excision of exon 5. b, RNAscope visualization of Chrnb2 mRNA in the DS and SNc in two representative mice injected with AAV-GFP or AAV-Cre-GFP. Chrnb2 mRNA puncta are missing in the DS injected with Cre while the SNc of the same animal shows no decrease in puncta density. c, Quantification of Chrnb2 mRNA puncta in the DS. t₍₄₎ = 7.824, p = 0.0014, two-tailed t test. d, Quantification of Chrnb2 mRNA puncta in the SNc. t₍₄₎ = 0.5321, p = 0.6228, two-tailed t test. e, Chrnb2 mRNA expression analyzed by qRT-PCR in striatal punches from AAV-expressing area of mice injected with AAV-GFP or AAV-Cre-GFP. t₍₁₁₎ = 16.95, p < 0.0001, two-tailed t test. f, Chrna4 mRNA expression analyzed by qRT-PCR in striatal punches from AAV-expressing area. t₍₁₁₎ = 0.5191, p = 0.6140, two-tailed t test. g, h, Chrnb2 mRNA expression analyzed by qRT-PCR in the cortex (g) and midbrain (h). Cortex: t₍₁₁₎ = 0.1788, p = 0.8613; midbrain: t₍₆₎ = 0.7645, p = 0.4736; two-tailed t test. i, Concentration of dopamine measured by ELISA in homogenates prepared from mice injected with AAV-GFP or AAV-Cre-GFP. t₍₁₄₎ = 1.106, p = 0.2874, two-tailed t test. Number of samples used for analysis is indicated in graphs. Data are individual values and mean ± SEM. str, Striatum; ctx, cortex; mdbr, midbrain.