• Figure 2.
    Download figureOpen in new tabFigure 2. Main stressors interfering with reliable measurement of social behavior in rodents (a) and their effects on social preference scores in C57BL/6 and BALB/c mice in the conventional three-chambered test (b–e) under low- and high-stress conditions.

    High-stress conditions differ from low-stress conditions in the intensity of light and subjects' and mouse social objects' habituation to the experimenter and the experimental environment (for a detailed protocol see 'Materials and methods'). (b) BALB/c mice showed social preference only in low-stress conditions (n = 17), and they avoided social interactions when tested in a typical experimental setting (n = 11). (c) In contrast, C57BL/6 mice displayed social preference in both stressful (n = 11) and stress-reducing (n = 38) conditions. Social preference was calculated as the time spent in the chamber containing the social object compared to the time spent in the chamber with a non-social object. (d,e) Under stress, both tested strains of mice showed reduced locomotor activity. Data are median values and error bars represent IQR (interquartile range), *p<0.05, **p<0.01, ***p<0.001 (Mann-Whitney U-test).

    DOI: http://dx.doi.org/10.7554/eLife.19532.010

    Figure 4.
    Download figureOpen in new tabFigure 4. Social impairment of Fmr1 knockout mice compared to wild-type control measured in Eco-HAB.

    Fmr1 knockouts, n = 22. Wild-type controls, n = 10. (a) Odor-based social preference in the Eco-HAB system defined as the increase in proportion of time spent in the compartment with social odor during the first hour after its presentation, divided by the proportion of time spent in the compartment with non-social stimulus. Histogram (b) shows the distribution of in-cohort sociability for all pairs of knockout and control animals. Data are mean values and error bars represent SEM, *p<0.05, **p<0.01, ***p<0.001 (Mann-Whitney U-test).

    DOI: http://dx.doi.org/10.7554/eLife.19532.018

    Figure 4—source data 1.Eco-HAB measured social approach and in-cohort sociability of Fmr1 knockouts and wild-type controls.

    The names of the Excel sheets refer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see 'Materials and methods').

    DOI: http://dx.doi.org/10.7554/eLife.19532.019

    Download source data [figure-4—source-data-1.media-5.xlsx]
    Figure 4—source data 2.Eco-HAB measured social approach and in-cohort sociability of Fmr1 knockouts and wild-type controls.

    The names of the Excel sheetsrefer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see 'Materials and methods').

    DOI: http://dx.doi.org/10.7554/eLife.19532.020

    Download source data [figure-4—source-data-2.media-6.xlsx]
    Figure 5.
    Download figureOpen in new tabFigure 5. Eco-HAB provides reproducible assessment of approach to social odor in group-housed mice.

    Individual results of approach to social odor for all cohorts of (a) valproate-treated (n = 20) and control (n = 18) BALB/c subjects (4 cohorts), (b) valproate-treated (n = 26) and control (n = 35) C57BL/6 mice (6 cohorts) and (c) Fmr1 knockouts (n = 22) and wild-type (n = 18) animals (5 cohorts). Each column represents one cohort of animals, while data points (dots and squares) represent scores of particular mice. Since the measure of approach to social odor is a proportion (for detailed description see 'Materials and methods'), which may take values from 0 to +∞, we present logarithmic data to depict reproducibility of social preference and social avoidance in an unbiased manner. All analyses, including statistical testing, were performed on raw data. Average results of these data are presented in Figures 3A,E and 4A, respectively.

    DOI: http://dx.doi.org/10.7554/eLife.19532.021

    Figure 5—source data 1.Eco-HAB measured social approach score for valproate-treated and control C57BL/6 and BALB/c mice and Fmr1 knockouts and wild-type controls.

    These data are identical to Figure 3A—source data 1, Figure 3E—source data 3, Figure 4A—source data 1 with respect to Figures 3A,E and 4A and are available as a separate file for the readers’ convenience.

    DOI: http://dx.doi.org/10.7554/eLife.19532.022

    Download source data [figure-5—source-data-1.media-7.xlsx]
    Figure 5—source data 2.Eco-HAB measured social approach score for valproate-treated and control C57BL/6 and BALB/c mice and Fmr1 knockouts and wild-type controls.

    These data are identical to Figure 3A—source data 1, Figure 3E—source data 3, Figure 4A—source data 1 with respect to Figures 3A,E and 4A and are available as a separate file for the readers’ convenience.

    DOI: http://dx.doi.org/10.7554/eLife.19532.023

    Download source data [figure-5—source-data-2.media-8.xlsx]
    Figure 5—source data 3Eco-HAB measured social approach score for valproate-treated and control C57BL/6 and BALB/c mice and Fmr1 knockouts and wild-type controls.

    These data are identical to Figure 3A—source data 1, Figure 3E—source data 3, Figure 4A—source data 1 with respect to Figures 3A,E and 4A and are available as a separate file for the readers’ convenience.

    DOI: http://dx.doi.org/10.7554/eLife.19532.024

    Download source data [figure-5—source-data-3media-9.xlsx]
    Figure 6.
    Download figureOpen in new tabFigure 6. Assessment of approach to social odor in Fmr1 knockouts and respective littermate controls performed in two different laboratories.

    (a, Fmr1 knockout n = 22, wild-type control n = 18) vs. (b, Fmr1 knockout n = 11, wild-type control n = 9). Regardless of experimental environment, evaluation carried out in Eco-HAB revealed comparable impairment in Fmr1 knockouts. Presented data are logarithmic values.

    DOI: http://dx.doi.org/10.7554/eLife.19532.025

    Figure 6—source data 1.We include source data for Figures 6, 7 and 8 concerning reproducibility results of both Eco-HAB measures.

    The names of the Excel sheets refer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see 'Materials and methods').

    DOI: http://dx.doi.org/10.7554/eLife.19532.026

    Download source data [figure-6—source-data-1.media-10.xlsx]
    Figure 6—source data 2.We include source data for Figures 6, 7 and 8 concerning reproducibility results of both Eco-HAB measures.

    The names of the Excel sheetsrefer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see Materials and methods).

    DOI: http://dx.doi.org/10.7554/eLife.19532.027

    Download source data [figure-6—source-data-2.media-11.xlsx]
    Figure 7.
    Download figureOpen in new tabFigure 7. Evaluation of in-cohort sociability in Fmr1 knockouts and wild-type littermate controls undertaken in two different laboratories.

    (a, Fmr1 knockout n = 22, wild-type control n = 18) vs. (b, Fmr1 knockout n = 11, wild-type control n = 9) – gives corresponding results. A histogram illustrating score of Fmr1 knockouts is shifted to the left as compared to that for wild-type control, signifying less time voluntarily spent together with other subjects within a tested cohort.

    DOI: http://dx.doi.org/10.7554/eLife.19532.028

    Figure 7—source data 1.We include source data for Figures 6, 7 and 8 concerning reproducibility results of both Eco-HAB measures.

    The names of the Excel sheetsrefer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see Materials and methods).

    DOI: http://dx.doi.org/10.7554/eLife.19532.029

    Download source data [figure-7—source-data-1.media-12.xlsx]
    Figure 7—source data 2.We include source data for Figures 6, 7 and 8 concerning reproducibility results of both Eco-HAB measures.

    The names of the Excel sheetsrefer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see Materials and methods).

    DOI: http://dx.doi.org/10.7554/eLife.19532.030

    Download source data [figure-7—source-data-2.media-13.xlsx]
    Figure 8.
    Download figureOpen in new tabFigure 8. Eco-HAB allows remarkably reproducible assessment of approach to social odor in both (a) wild-type mice (n = 9) and (b) Fmr1 knockouts (n = 11).

    Evaluation of social behavior of subjects was repeated twice in identical Eco-HAB experiments, separated by a 10-day period of regular housing. Each aligned dot and square encircled by an oval represent individual score of approach to social odor for each tested mouse, measured in two subsequent experimental repetitions. Dots are data, while the ovals serve to guide the eye. Data presented are logarithmic values.

    DOI: http://dx.doi.org/10.7554/eLife.19532.031

    Figure 8—source data 1.We include source data for Figures 6, 7 and 8 concerning reproducibility results of both Eco-HAB measures.

    The names of the Excel sheetsrefer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see Materials and methods).

    DOI: http://dx.doi.org/10.7554/eLife.19532.032

    Download source data [figure-8—source-data-1.media-14.xlsx]
    Figure 8—source data 2.We include source data for Figures 6, 7 and 8 concerning reproducibility results of both Eco-HAB measures.

    The names of the Excel sheetsrefer to corresponding figures and contain data used for analysis of the behavioral measures obtained by the implementation of Eco-HAB.py software (see Materials and methods).

    DOI: http://dx.doi.org/10.7554/eLife.19532.033

    Download source data [figure-8—source-data-2.media-15.xlsx]
  • Video 1. Top view of the working Eco-HAB.

    Flashing lights indicate activation of RFID antennas – sensors of the individual recognition system. The clip presents a 30 s period at the beginning of the adaptation phase, when animals are eagerly exploring new territory.

    DOI: http://dx.doi.org/10.7554/eLife.19532.009