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. 2024 Jan 19;29(5):1338–1349. doi: 10.1038/s41380-024-02413-y

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© The Author(s) 2024

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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Fig. 2 — a Diagram of the generation of MG-Bdnf-overexpressing mice. The tet-off system was activated in double transgenic mice (F1), which were obtained by crossing Iba1-tTA transgenic mice with Bdnf-tetO knock-in mice. Iba1-tTA(+)::Bdnf^(tetO/+) mice were the MG-Bdnf-overexpressing mice, and Bdnf^(tetO/+) mice were the control mice. b BDNF expression in microglia recovered from the cortex, as measured using ELISA. Iba1-tTA(+)::Bdnf^(tetO/+) mice had higher BDNF expression than Bdnf^(tetO/+) mice; this was normalized from day 5 using doxycycline (DOX) (The Kruskal-Wallis test, 9.086, p = 0.0118; n = 4 per group). c Schema of the behavioral experiments without doxycycline. The experiments were started at P61. d The Iba1-tTA(+)::Bdnf^(tetO/+) mice were less social in the three-chamber sociability test than the Bdnf^(tetO/+) mice. The Iba1-tTA(+)::Bdnf^(tetO/+) mice had a lower social interaction score than Bdnf^(tetO/+) mice (U = 47, p = 0.0226, Mann–Whitney U test, Bdnf^(tetO/+): n = 12, Iba1-tTA(+)::Bdnf^(tetO/+): n = 16) (left) and spent less time near novel mice (F_1,26(interaction) = 14.63, p = 0.0007, two-way ANOVA followed by Bonferroni post hoc test; Bonferroni post hoc analysis of Iba1-tTA(+)::Bdnf^(tetO/+) S vs. Bdnf^(tetO/+) S, p = 0.0001, Bdnf^(tetO/+): n = 12, Iba1-tTA(+)::Bdnf^(tetO/+): n = 16) (right). S, social; O, object. e Schematic of the Augmented Reality-based Long-term Animal Behavior Observing system (AR-LABO). Three novel C57BL/6 J mice were placed with a target mouse and observed for 1 h. f (Left) The cumulative number of approaches per 5-min bin for Bdnf^(tetO/+) and Iba1-tTA(+)::Bdnf^(tetO/+) mice. Iba1-tTA(+)::Bdnf^(tetO/+) mice accumulates significantly less cumulative approaches to other mice than Bdnf^(tetO/+) mice (two-way RM ANOVA, phenotype (control or Iba1-BDNF) × time (5 min bin) interaction F_(12,312) = 3.926, p < 0.0001; effect of phenotype F_(1,26) = 6.670, p = 0.0158; effect of time F_(12,312) = 39.31, p < 0.0001, Bdnf^(tetO/+): n = 15, Iba1-tTA(+)::Bdnf^(tetO/+): n = 13). (Right) Iba1-tTA(+)::Bdnf^(tetO/+) mice approached other mice less than the Bdnf^(tetO/+) mice (t₍₂₆₎ = 2.717, p = 0.0116, unpaired two-tailed Student’s t test, Bdnf^(tetO/+): n = 15, Iba1-tTA(+)::Bdnf^(tetO/+): n = 13). g The Iba1-tTA(+)::Bdnf^(tetO/+) mice were less approached by other mice than Bdnf^(tetO/+) mice (t₍₂₆₎ = 2.120, p = 0.0437, unpaired two-tailed Student’s t test, Bdnf^(tetO/+): n = 15, Iba1-tTA(+)::Bdnf^(tetO/+): n = 13). h No differences in activity were observed during AR-LABO between the Iba1-tTA(+)::Bdnf^(tetO/+) and Bdnf^(tetO/+) mice (t₍₂₆₎ = 0.8059, p = 0.4276, unpaired two-tailed Student’s t test, Bdnf^(tetO/+): n = 15, Iba1-tTA(+)::Bdnf^(tetO/+): n = 13). *p < 0.05, ***p < 0.001. Data are presented as the mean ± SEM. 2 M: two months of age, control: Bdnf^(tetO/+) mice, Iba1-Bdnf: Iba1-tTA(+)::Bdnf^(tetO/+) mice.