50454 aav8  (Addgene inc)

Journal: Experimental neurology

Article Title: Harnessing rAAV-retro for gene manipulations in multiple pathways that are interrupted after spinal cord injury

doi: 10.1016/j.expneurol.2021.113965

Figure Lengend Snippet: Use of intersectional genetic approaches. A: tdT expression reveals retrograde transduction of cortical motoneurons in one hemisphere with unilateral rAAV-retro/Cre injections at C5. Cortical motoneurons throughout the sensorimotor cortex, except for the hindlimb region, express tdT, whereas only a few neurons on the opposite side were transduced. B: Immunostaining of a different section for HA-citrine reveals the expression of the Cre-dependent hM3Dq with unilateral intra-cortical injections of AAV8-hM3Dq. C: Co-imaging for tdT (native fluorescence) reveals that hM3Dq is expressed exclusively in cortical motoneurons. D: immunostaining for c-fos 1 h following delivery of CNO in the mouse illustrated in A-C. E: Higher magnification views of c-fos labeling from the area indicated by rectangle in D; F: Co-imaging for tdT in the same section. G; Merge of D & E. (A-D scale bar: 500 μm; E-G: scale bar 250 μm).

Article Snippet: Intersectional genetic approach to selectively express DREADD in cortical motoneurons Rosa tdTomato mice, which express tdTomato upon Cre expression, received intra-spinal injections of rAAV-retro/Cre at cervical level 5 as above and unilateral intra-cortical injections of AAV carrying Cre-dependent hM3Dq (AAV8-hSyn-DIO-HA-hM3D(Gq)-IRES-mCitrine, Addgene Cat. No. 50454).

Techniques: Expressing, Transduction, Immunostaining, Imaging, Fluorescence, Labeling