Fig 1.
Flow chart of the test schedule.
+ Duration of resting [s], duration of locomotion [s], number of falls [n], and duration of episodes of rapid jerky dorsal-ventral head movements [s]; an.: anesthesia. This figure has been modified from Hohlbaum et al. [4, 32].
Table 1.
Phases of anesthesia.
Table 2.
Vital parameters.
Table 3.
Recovery period.
Fig 2.
Mouse Grimace Scale difference scores.
MGS, Mouse Grimace Scale; ♀: females; ♂: males. Data are presented as boxplot diagrams: the box represents the interquartile range (IQR), box edges are the 25th and 75th quartile. The whiskers represent values which are not greater than 1.5 × IQR. Dots are outliers with values between 1.5‒3.0 × IQR. Grey asterisks are outliers with values greater than 3.0 × IQR. Data were analyzed using Kruskal-Wallis-Test with post-hoc Dunn-Bonferroni test or, for sex differences, Mann-Whitney-U-Test ** p < 0.01 versus control). (A) Control ♀: n = 7, single anesthesia ♀: n = 13 at 150 min (n = 9 at day two; at this study day, four mice of the single anesthesia group were excluded from statistics due to technical malfunction of the camera), repeated anesthesia ♀: n = 13. (B) Control ♂: n = 6, single anesthesia ♂: n = 13, repeated anesthesia ♂: n = 12.
Table 4.
Nest building.
Table 5.
Home cage activity during the dark period (one day after the last anesthesia).
Table 6.
Physical fitness (two days after the last anesthesia).
Fig 3.
Latency to explore in the free exploratory paradigm for trait anxiety-related and exploratory behavior.
♀: females; ♂: males. Data are presented as boxplot diagrams: the box represents the interquartile range (IQR), box edges are the 25th and 75th quartile. The whiskers represent values which are not greater than 1.5 × IQR. Dots are outliers with values between 1.5 ‒ 3.0 × IQR. Grey asterisks are outliers with values greater than 3.0 × IQR. Data were analyzed using the Kruskal-Wallis-Test with post-hoc Dunn-Bonferroni test or, for sex differences, Mann-Whitney-U-Test (** p < 0.01 versus repeated anesthesia). (A) Control ♀: n = 7, single anesthesia ♀: n = 13, repeated anesthesia ♀: n = 13. (B) Control ♂: n = 6, single anesthesia ♂: n = 13, repeated anesthesia ♂: n = 12.
Table 7.
Food intake.
Fig 4.
Data are given as mean ± standard error. (A) Control ♀: n = 7, single anesthesia ♀: n = 13, repeated anesthesia ♀: n = 13. (B) Control ♂: n = 6, single anesthesia ♂: n = 13, repeated anesthesia ♂: n = 12. Data were analyzed using repeated measures ANOVA with group as between-subject factor (pairwise comparisons using Bonferroni correction: * p < 0.05, *** p < 0.001 versus day two; ** p < 0.01 versus day 14).
Table 8.
Fecal corticosterone metabolites (FCM) and hair corticosterone.