Effects of propofol and surgery on neuropathology and cognition in the 3xTgAD Alzheimer transgenic mouse model  F. Mardini, J.X. Tang, J.C. Li, M.J. Arroliga, R.G. Eckenhoff, M.F. Eckenhoff  British Journal of Anaesthesia  Volume 119, Issue 3, Pages 472-480 (September 2017) DOI: 10.1093/bja/aew397 Copyright © 2017 The Author(s) Terms and Conditions

Fig 1 Rodent learning and memory was tested using the Morris water maze. Spatial reference memory was tested in the 3xTgAD mice by measuring the escape latencies throughout 5 days at 3 weeks (a) and 15 weeks after treatment (b). No significant differences were detected at either time point using a two-way repeated-measures anova with Sidak's multiple comparisons test. (Path length data are shown in Supplementary Fig. 1a and b.) When the hidden submerged platform was removed for the probe test, the propofol and surgery group spent significantly less time in the target quadrant than the control mice (*P<0.01) at both 3 weeks (c) and 15 weeks (d) using a two-way anova with Sidak's multiple comparisons test (Tar, Target; Adj L, adjacent left; Opp, opposite; Adj R, adjacent right). There was no significant difference for the propofol-only group. (e) Spatial working memory testing was conducted during 10 days beginning at 16 weeks. The mean number of trials it took for each animal to reach criterion per platform was calculated; no difference was detected between the experimental groups and the control group using the Kruskal–Wallis anova test of means for non-parametric data with Dunn's multiple comparisons tests. (f) The swim speed was measured for all animals at each time point during the 60 s probe test; no significant differences in swim speeds were detected using a two-way anova with Sidak's multiple comparisons test. (g) Motor ability was also tested with the rotarod, and no significant differences in motor skill were detected between groups using a two-way anova with Sidak's multiple comparisons test. Data are expressed as means (95% confidence intervals). Animal numbers are as follows: at 3 weeks, n=12 for each group; at 15 weeks, n=8 control, n=12 propofol, and n=10 propofol and surgery. British Journal of Anaesthesia 2017 119, 472-480DOI: (10.1093/bja/aew397) Copyright © 2017 The Author(s) Terms and Conditions

Fig 2 Amyloid β was seen at 18 weeks post-operatively using immunohistochemistry with the 6E10 antibody and diaminobenzidine (brown). Representative examples of amyloid plaques (arrows) in the subiculum and CA1 region of the hippocampus are shown for the control (a), propofol (b), and propofol with surgery groups (c). Scale bar represents 50 µm. British Journal of Anaesthesia 2017 119, 472-480DOI: (10.1093/bja/aew397) Copyright © 2017 The Author(s) Terms and Conditions

Fig 3 Quantitative immunohistochemistry performed at 18 weeks. (a) The number of amyloid β plaques (6E10) per square millimetre of subiculum and CA1 was not significantly different in the propofol or propofol with surgery group compared with the control group. (b) In addition, the mean plaque area was not different between groups. (c) The number of phosphorylated tau (AT8)-positive cells in the hippocampal CA1 region and subiculum was not significantly different between groups. (d) Likewise, microglial activation (Iba-1) was not significantly different in the propofol or propofol with surgery group compared with the control group in the CA1 region of the hippocampus. Data were analysed with the Kruskal–Wallis anova test of means for non-parametric data with Dunn's multiple comparisons tests. The mean was calculated from five or six brain sections for each animal. Data are expressed as the mean (95% confidence interval), and n=5 animals for each group. British Journal of Anaesthesia 2017 119, 472-480DOI: (10.1093/bja/aew397) Copyright © 2017 The Author(s) Terms and Conditions

Fig 4 Brain sections were stained for phosphorylated tau immunoreactivity (AT8) in the subliculum and hippocampal CA1 area at 18 weeks. Representative examples of intraneuronal tau aggregates (arrows) in the subiculum and CA1 region of the hippocampus are shown for the control (a), propofol (b), and propofol with surgery groups (c). Scale bar represents 100 µm. British Journal of Anaesthesia 2017 119, 472-480DOI: (10.1093/bja/aew397) Copyright © 2017 The Author(s) Terms and Conditions

Fig 5 Microglial activation at 18 weeks was detected in the hippocampal CA1 region with Iba-1 immunohiostochemistry (arrows) in the three treatment groups: control (a), propofol (b), and propofol with surgery (c). Scale bar represents 100 µm. British Journal of Anaesthesia 2017 119, 472-480DOI: (10.1093/bja/aew397) Copyright © 2017 The Author(s) Terms and Conditions

Fig 6 Brain cytokine concentrations at 6 h postoperatively. Interleukin (IL)-10 (a) and IL-6 (b) concentrations were measured with enzyme-linked immunosorbent assays (ELISAs) in hippocampal tissue from 3xTgAD mice at 6 h. The ELISAs were repeated three times per animal. Data were analysed with the Kruskal–Wallis anova test of means for non-parametric data with Dunn's multiple comparisons tests. Data are expressed as the mean (95% confidence interval), and n=5 animals for each group. British Journal of Anaesthesia 2017 119, 472-480DOI: (10.1093/bja/aew397) Copyright © 2017 The Author(s) Terms and Conditions