Aav2 cre gfp

Adult male and female floxed Gabrd and Gabrg2 mice were stereotaxically injected with either AAV2-GFP or AAV2-Cre-GFP (Vector Biolabs) into the VTA using the following coordinates: A/P −3.6 mm, M/L ±0.5 mm, D/V 4.5 mm. This approach results in expression of GFP or Cre recombinase in a subset of neurons in the VTA.

Adult mice (Jackson Laboratory, Bar Harbor, ME) were handled in accordance with the Association for Research in Vision and Ophthalmology statement on the use of animals in research. All experimental protocols and the ethical care of the mice were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Wisconsin. Mice were housed in microisolator cages and kept on a 12-h light/dark cycle and maintained on a 4 % fat diet (8604 M/R; Harland Teklad, Madison, WI). Bax-deficient mice were generated from breeding Bax^+/− animals on a C57BL/6J background. Mice harboring LoxP sites flanking exons 3 and 4 in the Panx1 gene (Panx1^fl/fl), as previously described [78 (link)], were used to generate Panx1 total knockout mice and RGC conditional knockouts. For complete knockout mice, Panx1^fl/fl animals were crossed with transgenic mice carrying CRE recombinase under the control of the CMV immediate early promoter. For RGC-selective deletion of Panx1, Panx1^fl/fl mice received an intraocular injection of a replication-deficient AAV2 virus carrying a CRE expression cassette (AAV2-Cre/GFP, Vector Biolabs, Philadelphia, PA and University of North Carolina Viral Vector Core, Chapel Hill, NC), which transduces approximately 85 % of the RGCs with only minimal transduction of some Müller cells [79 (link), 80 (link)]. All genotypes were on the C57BL/6 background.