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. 2020 Nov 5;10:19178. doi: 10.1038/s41598-020-76183-2

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© The Author(s) 2020

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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Experimental timeline and representative image of DREADDs expression in the NAc. (a) Drd1a-iCre, Drd2-iCre, and Wt rats underwent 4-weeks of the intermittent access to the IA2BC20% alcohol-drinking paradigm prior to receiving intra-NAc infusions of either the stimulatory hM3D(G_q) or inhibitory hM4D(G_i) DREADDs viral constructs. Rats continued on the IA2BC20% alcohol-drinking paradigm for 3-weeks; during week-7, all rats received intraperitoneal (i.p.) injections of 5% DMSO 30-min prior to drinking; on the first session of week-8 (session 22), rats received either i.p. injections of CNO (3 mg/mL) or 5% DMSO and were allowed access to alcohol for 24-h. On session 23, rats received another injection of either CNO or DMSO 30-min prior to the introduction of alcohol and water bottles and were terminated 1-h after the start of drinking and 90-min after i.p. injection of CNO or DMSO. Brains were collected to examine cFos in the NAc 90-min after CNO administration and 1-h after alcohol intake. (b) Representative image of DREADDs virus expression within the nucleus accumbens (NAc) at 2× (left) and 4× objective. Images display expression of the DREADDs fluorescent reporter, mCherry, against DAPI counterstain.