Eco-HAB as a fully automated and ecologically relevant assessment of social impairments in mouse models of autism
- Nencki Institute of Experimental Biology of Polish Academy of Sciences, Poland
- Polish Academy of Sciences, Poland
- Warsaw University of Technology, Poland
- University of Zurich, Switzerland
- Kwazulu-Natal University Durban, Republic of South Africa
Figures
Figure 1 with 6 supplements
A schematic representation of Eco-HAB system and data processing.
Eco-HAB consists of four housing compartments (a), tube-shaped inter-territorial passages (b), radio-frequency identification antennas (c), and impassable, perforated partitions behind which social …
Figure 1—figure supplement 1
Block schematic diagram of customized electronic system for Eco-HAB.
https://doi.org/10.7554/eLife.19532.003
Figure 1—figure supplement 2
RFID antenna efficiency compared to video-based manual scoring.
We counted the number of RFID registrations per visually registered crossing under the antenna. The implemented system of coils recognizes subjects at a rate of at least one or more RFID …
Figure 1—figure supplement 3
Comparison of time (person-hours) needed for Eco-HAB testing versus three-chambered apparatus testing (stress reducing conditions) of a group of 12 mice.
For detailed description of behavioral protocols see 'Materials and methods'.
Figure 1—figure supplement 4
Eco-HAB measures in-cohort sociability in mice.
(a) Detailed data regarding quantity of time spent by each mouse with every other animal in the group can be obtained in Eco-HAB. (b) Based on simultaneous territory occupation for each individual, …
Figure 1—figure supplement 5
Eco-HAB allows for a detailed analysis of subjects' preference to spend time with another mouse from a tested cohort.
Examples show (a) a pair of mice spending most of their time together, regardless of their position within the territory, and (b) a pair of mice spending time mostly in different areas of Eco-HAB. …
Figure 1—figure supplement 6
Monitoring of subjects’ dispersal within Eco-HAB territory for exemplary cohorts of (a) C57BL/6 and (b) BALB/c mice.
Customized software provides easy access to data on the amount of time spent by a mouse in each Eco-HAB compartment. An example here shows mouse activity distribution in 12-hr dark phase during the …
Figure 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 …
Figure 3 with 2 supplements
Sociability measurements in Eco-HAB and three-chambered apparatus social approach test performed under stress reducing conditions.
Figure (a–d) depict tests involving C57BL/6 mice and (e–h) tests involving BALB/c mice. (a) and (e) show social approach in the Eco-HAB system defined as the increase in proportion of time spent in …
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Figure 3—source data 1
Eco-HAB measured social approach and in-cohort sociability of valproate-treated and control C57BL/6 and BALB/c mice.
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').
- https://doi.org/10.7554/eLife.19532.012
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Figure 3—source data 2
Eco-HAB measured social approach and in-cohort sociability of valproate-treated and control C57BL/6 and BALB/c mice.
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).
- https://doi.org/10.7554/eLife.19532.013
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Figure 3—source data 3
Eco-HAB measured social approach and in-cohort sociability of valproate-treated and control C57BL/6 and BALB/c mice.
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).
- https://doi.org/10.7554/eLife.19532.014
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Figure 3—source data 4
Eco-HAB measured social approach and in-cohort sociability of valproate-treated and control C57BL/6 and BALB/c mice.
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).
- https://doi.org/10.7554/eLife.19532.015
Figure 3—figure supplement 1
Three-chambered apparatus testing performed on group-housed valproate-treated C57BL/6 (VPA-treated n = 14, CTRL n = 38) (a) and BALB/c (n = VPA-treated n = 15, CTRL n = 17) (b) mice did not reveal any differences in sociability.
Data are represented as mean ± SEM.
Figure 3—figure supplement 2
Eco-HAB allows for long-term monitoring of responses to social stimuli.
We assessed subjects approach to social odor in four different periods, namely 30’, 1 hr, 2 hr, and 4 hr after the presentation of olfactory stimuli, in order to see how their behavior changes with …
Figure 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 …
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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').
- https://doi.org/10.7554/eLife.19532.019
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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').
- https://doi.org/10.7554/eLife.19532.020
Figure 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) …
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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.
- https://doi.org/10.7554/eLife.19532.022
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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.
- https://doi.org/10.7554/eLife.19532.023
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Figure 5—source data 3
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.
- https://doi.org/10.7554/eLife.19532.024
Figure 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 …
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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').
- https://doi.org/10.7554/eLife.19532.026
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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).
- https://doi.org/10.7554/eLife.19532.027
Figure 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 …
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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).
- https://doi.org/10.7554/eLife.19532.029
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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).
- https://doi.org/10.7554/eLife.19532.030
Figure 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 …
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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).
- https://doi.org/10.7554/eLife.19532.032
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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).
- https://doi.org/10.7554/eLife.19532.033
Figure 9 with 1 supplement
Tube dominance score of an animal does not correlate with overall activity in Eco-HAB apparatus.
Dominance is expressed as percentage of won encounters ('winning score percent') in the U-tube dominance test (see 'Materials and methods'). Activity in Eco-HAB is defined as the number of visits to …
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Figure 9—source data 1
Raw data from U-tube dominance test and Eco-HAB measured activity (number of visits to all compartments of the apparatus during 1st 12-hr period of adaptation).
Figure 9 depicts correlation between those two variables.
- https://doi.org/10.7554/eLife.19532.035
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Figure 9—source data 2
Raw data from U-tube dominance test and Eco-HAB measured activity (number of visits to all compartments of the apparatus during 1st 12 hr period of adaptation).
Figure 9 depicts correlation between those two variables.
- https://doi.org/10.7554/eLife.19532.036
Figure 9—figure supplement 1
Aggressive interactions during testing in Eco-HAB are rare regardless of the tested strain.
Number of episodes and duration of aggressive behaviors in VPA-treated and control BALB/c (a, b), VPA-treated and control C57BL/6 (c, d) and Fmr1 knockout and wild-type mice (e, f) during first 6 hr …
Videos
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 …
Additional files
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Supplementary file 1
Eco-HAB.py scripts with sample data enabling their execution.
- https://doi.org/10.7554/eLife.19532.038
