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. 2024 Mar 19;29(8):2496–2509. doi: 10.1038/s41380-024-02515-7

Fig. 2. Rfwd2+/− male mice showed a communication deficit in ultrasonic vocalization recording and impaired social behavior in the 3-chamber test.

Fig. 2

A Representative images of vocalizations recorded from Rfwd2+/− mice and WT littermates on postnatal (P) days 5, 7, and 9. Top, oscillogram signals. Bottom, corresponding spectrogram signals; all are 1.5 s long. B Quantification of the call rate revealed a decrease in the total number of calls made by Rfwd2+/− mice compared to WT littermates at P5, 7, and 9. C, D Mean (C) and total duration (D) of calls during the 5-min recording period at P5, P7 and P9 in Rfwd2+/− mice and WT littermates. E–G Mean number of calls emitted during the 5 min recording period at P5, P7, and P9. H Habituation phase: Rfwd2+/− and WT mice were allowed to explore the 3 chambers (center and the other two side chambers, containing two empty holding cells). I–K Rfwd2+/− and WT littermates spent similar amount of time in the two empty chambers and showed equal preference (E1/E2 ratio). L In the sociability test, Rfwd2+/− and WT mice were allowed to explore the 3 chambers, with one side chamber containing a novel mouse 1 (M1) and the other side chamber containing an empty (E) holding cell. N Rfwd2+/− mice and WT littermates displayed a reduced preference for the side chamber containing the empty holding cell than the chamber with the M1 mouse. O Rfwd2+/− mice showed a decreased preference (M1/E ratio) for mouse 1 compared to WT littermates. P In the social novelty test, a novel mouse (N) was placed in the opposite chamber, and M1 became a familiar mouse. Rfwd2+/− and WT mice were allowed to explore all the chambers. R Rfwd2+/− mice spent less time interacting with the novel mouse than the WT littermates and showed a decrease in preference (N/M1 ratio) for novel mice compared to WT littermates (S). Representative traces of ‘Empty 1-Empty 2’ (I) ‘Empty-Mouse 1’ (M) and ‘Novel mouse–Mouse 1’ (Q) for Rfwd2+/− mice and WT littermates. Data are shown as mean ± SEM. B–G Two-way repeated-measures ANOVA followed by Sidak’s test; (J, N, R), two-way ANOVA followed by Bonferroni’s test; (K, O, S), two-population Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001. n = 9, 10.