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. 2020 Jul 28;11:3764. doi: 10.1038/s41467-020-17543-4

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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 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. 1 — a Rats (n = 22) received Pavlovian conditioning sessions every other day in a distinct ‘alcohol context’ where a discrete, conditioned stimulus (CS) was paired with alcohol. On alternating days, rats were exposed to a different, ‘neutral context’ where a neutral stimulus (NS) was presented without alcohol. b Port-entries elicited by the CS (i.e., CS-triggered alcohol-seeking) were tested in the alcohol and neutral contexts by presenting the CS without alcohol. c At test, CS, but not PreCS, port-entries were elevated in the alcohol context compared to the neutral context [Context × Interval, F_(1,20) = 14.656, p = 0.001, η_p² = 0.423; t₍₂₁₎ = 3.736, p = 0.001, η_p² = 0.399]. d NonCS port-entries were unaffected by context [Context, F_(1,20) = 0.363, p = 0.554, η_p² = 0.018]. e We then conducted four additional tests in the alcohol and neutral contexts to determine if the impact of the alcohol context on CS-triggered alcohol-seeking was persistent. Normalized CS port-entries (CS minus PreCS; mean ± s.e.m.) decreased across repeated tests [Session, F_(3,30) = 18.435, p < 0.001, η_p² = 0.648], but were elevated in the alcohol context relative to the neutral context [Context, F_(1,10) = 30.671, p < 0.001, η_p² = 0.754; Context × Session, F_(3,30) = 5.885, p = 0.003, η_p² = 0.370]. Rats received the first test in the alcohol context (dashed line) or neutral context (solid line). f We then investigated the impact of context on relapse by presenting a drop of alcohol in the fluid port before and during the first CS presentation to reinstate CS port-entries. Half of the rats received the reinstatement test in the neutral context following their last extinction session in the alcohol context and vice versa. g Relative to an averaged baseline of the last two extinction sessions, normalized CS port-entries were reinstated in both contexts [Phase, F_(1,20) = 108.159, p < 0.001, η_p² = 0.844], but to a greater extent in the alcohol context than in the neutral context [Phase × Reinstatement Context, F_(1,20) = 6.037, p = 0.023, η_p² = 0.232; t₍₂₀₎ = 2.112, p = 0.047, η_p² = 0.182]. Averaged data are mean ± s.e.m. with data from individual rats overlaid on the bar graphs. Data were analysed using RM ANOVA (c, d, e, g) and Bonferroni-corrected t tests (c, g). All statistical tests were two-sided. Raw data are available as a supplementary source data file.