Article Figures & Data
Figures
- Figure 1.
PS19 mice have decreased sleep that worsens with age. A, Experimental design. B, Twenty-four hours of tracing the average hourly sleep of female WT (blue line) PS19 (pink line) mice at 3 months (prepathology), 6 months (early-phase), 9 months (symptomatic-phase), and 11 months (end-stage). The gray bars in sleep traces indicate a dark phase. C, D, Quantification of average hourly sleep (C) and sleep bout length in seconds (D). Data separated into 12 h of dark and light phases. E, Twenty-four hours of tracing the average hourly sleep of male WT (blue line) and PS19 (pink line) mice at 3, 6, 9, and 11 months. F, G, Quantification of average hourly sleep (F) and sleep bout length in seconds (G). Data separated into 12 h of dark and light phases. N = 5–17/age/sex/genotype. *p < 0.05, **p < 0.01, ***p < 0.001. Unpaired two-tailed Student's t test. Error bars indicate ± SEM. Estimated NREM and REM sleep % time in state and bout lengths are presented in Extended Data Figure 1-1 (females) and Extended Data Figure 1-2 (males).
- Figure 2.
Chronic sleep disruption accelerates cognitive decline in males. A, Experimental design. B, Males, Acquisition of spatial learning. Escape latencies during training in control and CSD treated mice. C, Males, Spatial memory retention during 1 min of probe trial, time in target or opposite quadrant in control and CSD treated mice. Target indicates the quadrant where the platform had been located, versus the opposite quadrant. D, Females, Acquisition of spatial learning. Escape latencies during training in control and CSD treated mice. E, Females, Spatial memory retention during 1 min of probe trial, time in target or opposite quadrant in control and CSD treated mice. N = 8–17 per group (sex, genotype, treatment). B, D, Data are means(±SEM) of four trials per day. C, E, Data are means (+SEM). *p < 0.05, **p < 0.01.
- Figure 3.
Tau in the cortex postsynaptic density (PSD). A, Subcellular fractionation technique to isolate PSD. The red line indicates the synaptosome fraction used to further isolate the PSD. B, C, Western blot analysis of cortex PSD samples showing soluble phosphorylated Tau (AT8) accumulation with age in PS19 and WT females (B) and males (C). Western blots also include total Tau and PSD95, a protein enriched in the PSD.
- Figure 4.
Decreased sleep amount in PS19 Tau tg mice is not predictive of AT8-Tau pathology in the cortex. A, Experimental design. B, Western blot analysis of AT8, AT180, AT100, pS396, total Tau, and PSD95 in 6-month-old (early-phase) PS19 females. C, D, Correlation of AT8-Tau hyperphosphorylation expression in the cortex of PS19 females with average dark-phase hourly sleep (C) or sleep bout length in seconds (D). Sleep data were separated into 12 h of dark and light phases. Dark-phase sleep is represented here. N = 16 PS19 females. E, Western blot analysis of AT8, AT180, AT100, pS396, total Tau, and PSD95 in 6-month-old PS19 males. F, G, Correlation of AT8-Tau hyperphosphorylation expression in the cortex of PS19 males with average dark-phase hourly sleep (F) or sleep bout length in seconds (G). Sleep data were separated into 12 h of dark and light phases. Dark-phase sleep is represented here. N = 11. All antibodies normalized to loading control. No significance (Pearson’s correlation). See Extended Data Figure 4-1 for further correlation analysis between sleep metrics and phosphorylated Tau.
- Figure 5.
Synaptic Tau is not increased in PS19 mice after acute sleep deprivation in the cortex and hippocampus. A, Experimental design. B, Western blot analysis of AT8, AT180, AT100, pS396, Tau1, and total Tau cortical expression in 6-month-old (early pathology) PS19 females and males. C, Quantification of cortical synaptic Tau proteins in PS19 females and males. N = 3 control; 3 CSD per sex. D, Western blot analysis of AT8, AT180, AT100, pS396, Tau1, and total Tau hippocampal expression in 6-month-old (early pathology) PS19 females and males. E, Western blot analysis of AT8, AT180, AT100, pS396, Tau1, and total Tau hippocampal expression in 6-month-old (early pathology) PS19 females and males. N = 3 control; 3 SD per sex. All antibodies normalized to loading control and then normalized to the control group. Unpaired two-tailed Student's t test between the control and treatment groups. Error bars indicate mean ± SEM.
- Figure 6.
Synaptic Tau is not increased in PS19 mice after chronic sleep disruption in the cortex and hippocampus. A, Experimental design. B, Western blot analysis of AT8, AT180, AT100, pS396, Tau1, and total Tau in cortical PSD fractions of 6-month-old (early pathology) PS19 females and males. C, Quantification of cortical synaptic Tau proteins in PS19 females and males. N = 5 control; 5 CSD females; N = 5 control; 6 CSD males. D, Western blot analysis of AT8, AT180, AT100, pS396, Tau1, and total Tau in the hippocampi of 6-month-old PS19 females and males. E, Quantification of hippocampal Tau proteins in PS19 females and males. N = 5 control; 5 CSD females; N = 4 control; 6 CSD males. All antibodies normalized to loading control and then normalized to the control group. Unpaired two-tailed Student's t test between the control and treatment groups. Error bars indicate mean ± SEM.
- Figure 7.
Chronic sleep disruption is a driver of Tau pathology in LC. A, Histology of locus ceruleus sections from PS19 mice under control or following 30 d of CSD treatment. Slices stained with TH to mark the noradrenergic neurons and AT8 to mark the Tau pathology. Example images from female PS19 mice. B, % area positive for AT8 in females, males, or combined. No AT8 signal was detected in WT littermates; therefore, statistical analysis was only conducted comparing control and CSD treatment in PS19 animals. CSD drove a significant increase in AT8 % area compared with control treatment in PS19 females or combined sexes. PS19 males showed a trend to increase. *p < 0.05 unpaired two-tailed Student's t test between the control and treatment groups for PS19 genotype. Error bars indicate mean ± SEM.
- Figure 8.
Chronic sleep disruption leads to changes in hippocampal synaptic protein expression in PS19 females but not males. A, B, Western blot analysis of hippocampal synaptic receptor expression in 6-month-old PS19 females (A) and males (B). C, Quantification of hippocampal protein expression in PS19 females. N = 5 control; 5 CSD. D, Quantification of hippocampal protein expression in PS19 males. N = 4 control; 6 CSD. All antibodies normalized to loading control and then normalized to the control group. Unpaired two-tailed Student's t test between the control and treatment groups for each genotype. *p < 0.05, **p < 0.01, ***p < 0.001. Error bars indicate mean ± SEM. E, F, Western blot analysis of cortical synaptic protein expression in 6-month-old PS19 females (E) and males (F). G, Quantification of cortical synaptic protein expression in PS19 females. N = 5 control; 5 CSD. H, Quantification of cortical synaptic protein expression in PS19 males. N = 4 control; 6 CSD. All antibodies normalized to loading control and then normalized to the control group. Unpaired two-tailed Student's t test between the control and treatment group for each genotype. Error bars indicate mean ± SEM.
Tables
Antibody Species Company Concentration Anti-AT8 Mouse Thermo Fisher Scientific 1:1,000 Anti-AT180 Mouse Thermo Fisher Scientific 1:1,000 Anti-AT100 Mouse Thermo Fisher Scientific 1:1,000 Anti-pS396 Rabbit Thermo Fisher Scientific 1:1,000 Anti-Tau1 Mouse EMD Millipore 1:1,000 Anti-K9 Rabbit Agilent Technologies 1:5,000 Anti-PSD95 Mouse NeuroMab 75-028 1:1,000,000 Anti-GluA1 Mouse NeuroMab 75-327 1:1,000 Anti-pS845 Rabbit EMD Millipore 1:1,000 Anti-pS831 Rabbit EMD Millipore 1:1,000 Anti-GluA2 Mouse EMD Millipore 1:1,000 Anti-GluA3 Rabbit Cell Signaling Technology 1:1,000 Anti-Arc Mouse Synaptic Systems 1:1,000 Anti-GluN2A Rabbit Cell Signaling 1:1,000 Anti-GluN2B Rabbit Cell Signaling Technology 1:1,000 Anti-GluN1 Rabbit Cell Signaling Technology 1:1,000 Anti-Hsc70 Mouse EMD Millipore 1:1,000
Table 1-1
Summary sleep measures from Figure 1. Sleep amount: % of time spent sleeping in the light or dark phase. Average sleep bout length (s) in light and dark phase. Download Table 1-1, XLSX file.
Figure 1-1
PS19 female mice exhibit progressive decrease in REM and NREM sleep. (A) 24hr trace of NREM sleep in female WT (blue line) PS19 (pink line) mice at 3, 6, 9, 11 months. Grey bars in sleep traces indicate dark phase. (B and C) Quantification of average hourly NREM sleep amount (B) and NREM sleep bout length in seconds (C). (D) 24hr trace of REM sleep in female WT (blue line) PS19 (pink line) mice at 3, 6, 9, 11 months. Grey bars in sleep traces indicate dark phase. (E and F) Quantification of average hourly REM sleep amount (E) and REM sleep bout length in seconds (E). Data separated into 12hrs of dark and light phases. N = 5-17/age/genotype. *p < 0.05, **p < 0.01, ***p < 0.001 Unpaired two-tailed student’s t-test. Error bars indicate ± SEM. Download Figure 1-1, TIF file.
Figure 1-2
PS19 male mice exhibit progressive decrease in REM and NREM sleep. (A) 24hr trace of NREM sleep in male WT (blue line) PS19 (pink line) mice at 3, 6, 9, 11 months. Grey bars in sleep traces indicate dark phase. (B and C) Quantification of average hourly NREM sleep amount (B) and NREM sleep bout length in seconds (C). (D) 24hr trace of REM sleep in male WT (blue line) PS19 (pink line) mice at 3, 6, 9, 11 months. Grey bars in sleep traces indicate dark phase. (E and F) Quantification of average hourly REM sleep amount (E) and REM sleep bout length in seconds (E). Data separated into 12hrs of dark and light phases. N = 5-17/age/genotype. *p < 0.05, **p < 0.01, ***p < 0.001 Unpaired two-tailed student’s t-test. Error bars indicate ± SEM. Download Figure 1-2, TIF file.
Table 1-2
Summary of estimated NREM and REM sleep measures from extended data Figures 1-1 and 1-2. Estimated NREM/REM Sleep amount: % of time spent sleeping in the light or dark phase. Average estimated NREM/REM sleep bout length (s) in light and dark phase. Download Table 1-2, XLSX file.
Figure 4-1
Decreased sleep amount in PS19 Tau tg mice is not predicative of AT8 Tau pathology in the cortex (continued). Western blots analysis of AT180, AT100, pS396 (see Figure 3) correlated to dark phase sleep measures in 6-month (early phase) PS19 females and males. (A and B) Correlation analysis of AT180, AT100, pS396 Tau pathology expression in the cortex of PS19 females with average dark phase hourly sleep (A) or sleep bout length in seconds (B). N = 16 PS19 females. (C and D) Correlation analysis of AT180, AT100, pS396 Tau pathology expression in the cortex of PS19 males with average dark phase hourly sleep (C) or sleep bout length in seconds (D). N = 11 PS19 males. All antibodies normalized to loading control. No significance (Pearson correlation). Download Figure 4-1, TIF file.
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