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. 2017 Jul 5;7:4739. doi: 10.1038/s41598-017-04972-3

Figure 1.

Figure 1

Vgat-Cre + SDH interneuron-specific hM4Di expression. (a) Schematic illustration of the FLEX switch system. AAV2/9 vector carrying EF1α and an inverted hM4Di-P2A-mCherry sequence flanked by loxP and lox2272 on both sides. Cre-dependent genetic inversion switch of the hM4Di-P2A-mCherry sequence occurs in Cre-expressing GABAergic SDH interneurons of Vgat-Cre mice. (b) mCherry expression in the SDH of mice 4 weeks after intra-SDH injection of AAV2/9-FLEX-hM4Di-P2A-mCherry. (c–e) Immunohistochemical identification of mCherry-expressing cells using cell-type specific markers (green; (c) PAX2; (d) Iba1; (e) GFAP). Arrowheads indicated mCherry+ cells with PAX2 immunofluorescence. (f) Photomicrographs of mCherry+ SDH neurons (upper, epifluorescence; lower, Nomarski optics) in spinal cord slices taken from Vgat-Cre;AAV-hM4DiFLEX mice. (g) Representative firing pattern of an mCherry+ SDH neuron in a spinal cord slice. (h) Representative traces of the effect of CNO (10 μM) on mCherry+ (left) and mCherryneg (right) SDH neurons. (i) Quantification of membrane potentials in mCherry+ SDH neurons before (Pre) and after application of CNO (n = 5, **P < 0.01).