addgene aavretro pgk cre  (Addgene inc)

Journal: The Journal of Comparative Neurology

Article Title: Local targets of T‐stellate cells in the ventral cochlear nucleus

doi: 10.1002/cne.25378

Figure Lengend Snippet: Labeling of T‐stellate cells via viral injection. (a) Schematic of the experimental approach for identifying T‐stellate cells. The rAAV2‐retro‐PKG‐Cre vector was injected into the inferior colliculus (IC) of Ai14 mice to retrogradely infect T‐stellate cells and allow the selective expression of tdTomato in infected neurons. (b) A small volume of injection (plus sign) results in a banding pattern of tdTomato expression (magenta) in the injected IC (outlined with dashed lines). DNA was counterstained with DAPI (blue). The stitched images were placed on black backgrounds for the purpose of visualizing the images all at the same orientation as the schematic in (a). (c) Distribution of tdTomato‐labeled T‐stellate cells in the contralateral cochlear nucleus (CN) from one experiment. Four coronal sections spanning the CN posterior‐anterior axis showing tdTomato‐labeled T‐stellate cells (magenta) distributed along a band within both the posterior ventral CN (pVCN) and anterior VCN (outlined with white dashed lines). DNA was counterstained with DAPI (blue). Axons of T‐stellate cells exit the CN via the ventral acoustic stria (asterisks). Scale bars in (b) = 500 μm, (c) = 200 μm. CB, cerebellum

Article Snippet: A total of 60–80 nl of AAV vector (rAAV2‐retro‐pkg‐Cre, Addgene (Watertown, MA) 24593‐AAVrg; Tervo et al., ) or 400 nl of BDA (3000 MW, Invitrogen, 10% in sodium citrate‐HCl, pH = 3) was loaded into a glass pipette with a tip diameter of 10–15 μm, and pressure was injected into the IC at a rate of 1 nl/s.

Techniques: Labeling, Injection, Plasmid Preparation, Expressing, Infection