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. 2019 Jul 12;10:3075. doi: 10.1038/s41467-019-10770-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — GFP Immunolabeling following transfection with the AAV8/CAG-ArchT-GFP construct. GFP staining (dark black/brown, shown for case F1115) was used as a reporter of ArchT expression. a GFP immunoreactivity following AAV8/CAG-ArchT-GFP injections in A1 and GFP-labeled axons and terminals in the medial geniculate body (MGB). b GFP-labeled axons and terminals in the inferior colliculus (IC). c–f High-power views. GFP-immunopositive (c) and NeuN-immunopositive (d) neurons at the location of the injection site in the cortex and, in the same animal, GFP labeling in fibers and terminals in the MGB (e) and IC (f). ArchT expression was identified on the basis of GFP immunoreactivity in the auditory cortex and compared with the distribution of NeuN-immunopositive neurons in order to estimate transfection rates for the two different constructs in ferrets used for behavioral testing (30 months after injections) and those used only for anatomy or recordings (4 weeks after injections). Stereological estimations made using the optical fractionator probe (n = 5 cases, 3 using CAG and 2 using CaMKII promoters) revealed that when AAV8/CAG-ArchT-GFP was used, 84.4 ± 14.8% (mean ± SD) of the neurons in the injection site were transfected, whereas, with AAV8/CaMKII-ArchT-GFP, the transfection rate was 60.03 ± 14.8% (mean ± SD). Other abbreviations: A1 primary auditory cortex, D dorsal, EG ectosylvian gyrus, LGN lateral geniculate nucleus, M medial, SSG suprasylvian gyrus, sss suprasylvian sulcus, V ventral. Calibration bars, 1 mm in (a) and (b), 50 µm in (d). Orientation and calibration bars in (d) apply to (c, e, f)