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Fig. 3 | Molecular Brain

Fig. 3

From: GABAergic neuron-specific whole-brain transduction by AAV-PHP.B incorporated with a new GAD65 promoter

Fig. 3

Specific and efficient transduction of cortical GABAergic interneurons by the mGAD65 promoter. VGAT-tdTomato mice that express tdTomato specifically in GABAergic neurons received intravenous (i.v.) injections of blood–brain barrier (BBB)-penetrating AAV-PHP.B expressing GFP under the control of the mGAD65 promoter or mDlx enhancer. The motor cortex was observed 3 weeks after the viral injection by confocal microscopy. a-d Low (a, b) and high (c, d) magnified images of native GFP and tdTomato. Arrows indicate cells double-labeled with GFP and tdTomato (GABAergic interneurons). Red arrowheads show GFP expression in tdTomato-negative cells (non-GABAergic cells). Scale bars; 50 μm (a, b) and 20 μm (c, d). e Graph showing highly specific transduction of cortical interneurons by the mGAD65 promoter. The specificity was determined by the ratio of double-labeled cells to total GFP ( +) cells. The value obtained from each mouse was plotted on the bar. For the mGAD65 promoter, we assessed 1553 GFP ( +) cells in 6 sections from 6 animals, and we found that 1483 cells were co-labeled with tdTomato (95.5 ± 0.7%). For the mDlx enhancer, we examined 1161 GFP ( +) cells in 5 sections from 5 animals, and we found that 1025 cells were co-labeled with tdTomato (88.3 ± 1.3%). ***p = 0.0007 by unpaired t-test. f Graph showing transduction efficiency of cortical interneurons by the mGAD65 promoter, in comparison to the mDlx enhancer. Transduction efficiency was determined by the proportion of GFP ( +) cells in tdTomato ( +) cells. For the mGAD65 promoter, we counted 2132 tdTomato ( +) cells in 6 sections from 6 animals, in which 1268 cells were GFP positive (60.1 ± 2.1%). For the mDlx enhancer, we observed 1503 tdTomato ( +) cells in 5 sections from 5 animals, in which 850 cells were GFP positive (56.5 ± 6.0%). p = 0.55 by unpaired t-test

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