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. 2018 Jun 15;43(10):1989–1999. doi: 10.1038/s41386-018-0119-4

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© The Author(s) 2018

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. 2 — Addicted mice have more silent synapses in DG during cue relapse than non-addict mice. a Experimental timeline and IntelliCage setups for alcohol drinking and alcohol-naive mice. Mice went through tests measuring addiction-related behaviors: motivation, persistence, withdrawal, cue relapse, and alcohol relapse spaced by periods of free access to alcohol, to identify addict and non-addict animals. b Addicts had significantly higher addiction score than non-addicts (t(27) = 6.844, p < 0.0001). c Mice performance during the training. (i) Addict and non-addict mice did not differ in nose-poke activity during the cage adaptation (CA) and initial 30 days of free access to alcohol (30d). (ii) They also did not differ in alcohol consumption during initiation of alcohol consumption (4 and 8%). Later (90d) addicts, as compared with non-addict mice, (i) performed more nosepokes to cage corners, and (ii) consumed more alcohol. They also (iii) reached higher breakpoint during Motivation tests, (iv) showed higher increase of reward nosepokes during non-rewarded (nR) phases as compared to rewarded (R) phases during persistence tests; (v) performed more nosepokes to the reward corner during withdrawal; (vi) performed more nosepokes to the reward corner during presentation of alcohol predicting cue; (vii) drank more alcohol during 12 h of relapse (test) as compared with non-addicts, and to the last 12 h of the last day of free alcohol access period (“0”). d Electrophysiological analysis of the granule cells in dorsal DG. (i) Experimental timelines and cage setups for alcohol drinking and alcohol-naive mice. Mice were killed during period of free access to alcohol (day 115), withdrawal (day 122) or cue relapse (+90’). (ii) Recording electrode in dorsal DG. Stimulating electrode was in perforant path. (iii) Example EPSCs (successes and failures) elicited by minimal stimulations at +45 mV (top) and −60 mV (bottom). Frequency of successes and failures was used to calculated % of silent synapses (see Materials and Methods for details). e Electrophysiological analysis. Trial plots of EPSCs elicited by minimal stimulations at +45 and −60 mV from alcohol-naive, non-addict and addict mice killed (i) during free alcohol drinking, (ii) withdrawal, and (iii) cue relapse, and mice drinking water. (iv) Frequency of silent synapses was increased in alcohol-drinking mice (both addict and non-addict) as compared to alcohol-naive animals during free access to alcohol, and addict mice as compared to non-addict and alcohol-naive mice during cue relapse *p < 0.05, **p < 0.01 by Tukey’s multiple comparisons test