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. 2019 Oct 14;9:14768. doi: 10.1038/s41598-019-51201-0

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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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AAV viral vectors are highly specific for Cre-expressing kisspeptin neurons. (A) Representative confocal images of the rostral (rARC), middle (mARC) and caudal (cARC) arcuate nucleus containing Kiss1-Cre cells expressing tdTomato (i), cells immunoreactive for GFP (AAV-TVA/GFP) and detected using a secondary antibody at 650 nm pseudocolored in green (ii) and merged images (iii) with insets illustrating the high degree of colocalization between channels. Arrow points to a rare non-kisspeptin/tdTomato cell transfected with AAV-TVA/GFP. (B) Histogram depicting the percentage of r-cARC kisspeptin neurons transfected by AAV-TVA/GFP, as shown by the mean ± SEM percentage of Kiss1-Cre/tdTomato positive cells colocalized with AAV-GFP. (C) Histogram depicting the high specificity of AAV-TVA/GFP for Cre, as shown by the mean ± SEM percentage of GFP-immunoreactive cells (AAV-GFP) colocalized with Kiss1-Cre/tdTomato-positive neurons in the r-cARC. No significant differences in the specificity of viruses or the percentage of tdTomato cells transfected were detected between male and female mice in the r-cARC (Student’s t-test).