Fig 1.
Frequency of observed behavior decreases during habituation.
(A) Distance travelled [cm] and (B) average activity [%] are depicted. Values are presented as exact value per each animal (n = 38 C57BL/6J, n = 15 BALB/cJ and n = 15 129S1/SvImJ male mice). The Friedman test with Dunn’s multiple comparisons test was used for statistical evaluation (n.s. = not significant (A) p-value = 0.7119, (B) p-value > 0.9999, **** p-value < 0.0001 (A, B).
Table 1.
Variances of distance travelled and average activity within the habituation period.
Table 2.
Repeatability values with multiple grouping factors: animal ID, strain and experiment.
Fig 2.
Animal ID and strain repeatability increase during habituation.
(A, B) Calculated animal ID repeatability value and (C, D) strain repeatability value for the factors (A, C) distance travelled and (B, D) average activity were presented. Each repeatability value (R, black points) was calculated over three adjacent days resulting in five groupings (n = 38 C57BL/6J, n = 15 BALB/cJ and n = 15 129S1/SvImJ male mice). Estimation of repeatability was conducted with a linear mixed-effect model based on Gaussian distribution. The [2.5%, 97.5%] confidence intervals (CI) were displayed in green, resulting from 500 bootstrapping runs and 100 permutations.
Fig 3.
Animal ID repeatability adjusted for strain increase during habituation.
Calculated animal ID repeatability values adjusted for the fixed factor strain were presented for (A) distance travelled and (B) average activity. Each repeatability value (R, black points) was calculated over three adjacent days resulting in five groupings (n = 38 C57BL/6J, n = 15 BALB/cJ and n = 15 129S1/SvImJ male mice). Estimation of repeatability was conducted with a linear mixed-effect model for locomotion as well as activity and with a generalized linear mixed-effect model based on Gaussian distribution for rearing and sniffing behavior. The [2.5%, 97.5%] confidence intervals (CI) were displayed in green, resulting from 500 bootstrapping runs and 100 permutations.
Fig 4.
Strategy for the implementation of behavioral experiments.
This basic strategy can be applied to all experiments with laboratory animals to determine the optimal starting point of your experiment which needs a familiar environment.