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. 2021 Apr 22;11:8788. doi: 10.1038/s41598-021-88200-z

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Virally mediated NMDAR ablation. (A) Illustration of target region of the medial PFC (blue), with the PrL region in darker shade of blue, in a saggital cut through the mouse brain. Vertical lines indicate infusion sites for broader (dotted, dorsal PFC) and more narrow (solid, PrL) targeting. (B, C) Examples of staggered coronal slices of a mouse brain transfected with rAAV8-CamKII $α$ -GFP-Cre (green) either with broader (B) or more narrow (C) targeting (see Methods). Blue, DAPI-stain; green, native GFP-fluorescence. Scale bar, 1 mm. Numbers state the distance from Bregma (as shown in A). (D) Microscopic image showing expression of GFP (green) and GluN1 (magenta) of a brain slice (at the PFC/M2 border) of an fGrin1 mouse brain slice transfected with a GFP-Cre vector; note the ample expression of GluN1-positive cells outside the transfected region. (E) Closer view onto a section of GFP-Cre-transfected (green) PFC double-stained against GluN1 (magenta) and Gad67 (cyan). Italic G are placed to the bottom left of Gad67-positive cells. (F) Section of a PFC of a GFP-transfected control mouse, stained against GluN1. Note that GFP-expression from the control vector is cytosolic (F), but GFP is localized to the nucleus if fused to Cre (E). GFP-positive cells are labelled by white asterisks. Scale bar, 50 $μ$ m (D-F). (G) Bar-graphs showing average share of GFP-positive cells that were also positive for GluN1 in the indicated groups and at the stated time-points after transfection. (H) Average share of Gad67-positive cells that were also positive for GluN1 (left) or GFP (right) in the indicated groups (the time points 3–4 wks and 7 wks after transfection were merged for this analysis; no Gad67-positive cells were GFP-positive). Stated N-numbers in each bar of (G-H) refer to analysed fields of view (top) obtained from the number of mice stated in brackets below. ***P < 0.001, MWU-test. (I) Order and approximate duration of behavioural assays (black) conducted in this study after the initial surgery (grey).