Eco-HAB as a fully automated and ecologically relevant assessment of social impairments in mouse models of autism

  1. Alicja Puścian
  2. Szymon Łęski
  3. Grzegorz Kasprowicz
  4. Maciej Winiarski
  5. Joanna Borowska
  6. Tomasz Nikolaev
  7. Paweł M Boguszewski
  8. Hans-Peter Lipp
  9. Ewelina Knapska  Is a corresponding author
  1. Nencki Institute of Experimental Biology of Polish Academy of Sciences, Poland
  2. Polish Academy of Sciences, Poland
  3. Warsaw University of Technology, Poland
  4. University of Zurich, Switzerland
  5. Kwazulu-Natal University Durban, Republic of South Africa
9 figures, 1 video and 1 additional file

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 …

https://doi.org/10.7554/eLife.19532.002
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 …

https://doi.org/10.7554/eLife.19532.004
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'.

https://doi.org/10.7554/eLife.19532.005
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, …

https://doi.org/10.7554/eLife.19532.006
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. …

https://doi.org/10.7554/eLife.19532.007
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 …

https://doi.org/10.7554/eLife.19532.008
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 …

https://doi.org/10.7554/eLife.19532.010
Figure 3 with 2 supplements
Sociability measurements in Eco-HAB and three-chambered apparatus social approach test performed under stress reducing conditions.

Figure (ad) depict tests involving C57BL/6 mice and (eh) 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 …

https://doi.org/10.7554/eLife.19532.011
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
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
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
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.

https://doi.org/10.7554/eLife.19532.016
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 …

https://doi.org/10.7554/eLife.19532.017
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 …

https://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').

https://doi.org/10.7554/eLife.19532.019
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
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) …

https://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.

https://doi.org/10.7554/eLife.19532.022
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
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
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 …

https://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').

https://doi.org/10.7554/eLife.19532.026
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
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 …

https://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).

https://doi.org/10.7554/eLife.19532.029
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
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 …

https://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).

https://doi.org/10.7554/eLife.19532.032
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 …

https://doi.org/10.7554/eLife.19532.034
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
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 …

https://doi.org/10.7554/eLife.19532.037

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 …

https://doi.org/10.7554/eLife.19532.009

Additional files

Supplementary file 1

Eco-HAB.py scripts with sample data enabling their execution.

https://doi.org/10.7554/eLife.19532.038

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