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. 2021 Apr 29;11:9335. doi: 10.1038/s41598-021-88599-5

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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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Lack of anxiolytic effect of diazepam in restraint-stressed animals. (A) (Left) Cumulative number of entries in the open arms of the maze along the 10-min session. (Middle and right) Mean (± SD) number of open arm entries after 5 (middle) and 10 min (right). White circles show data for individual animals. Animals were subjected to 1-h restraint-stress and then injected i.p. with vehicle or three doses of diazepam (0.5, 1.0 and 2.0 mg/kg) thirty minutes prior to behavior testing, as labeled. (B–D) Panels show the same as in (A), but for the percentage of the time spent in the open arms (B), and the total time animals spent moving (C) or in stretch-attend posture (D). Diazepam exhibited no statistically significant anxiolytic effect in this protocol, nor influenced locomotor activity. As in non-restrained animals (Fig. 1D), diazepam dose-dependently reduced the number of risk assessment behavior shown as reduced time in stretch-attend posture. *p < 0.01, **p < 0.001, ***p < 0.0001, one-way ANOVA followed by Tukey’s post hoc test. In A and B, outliers above the y-axis limit are not shown.