Differential effects of light and feeding on circadian organization of peripheral clocks in a forebrain Bmal1 mutant
- University of Texas Southwestern Medical Center, United States
- Northwestern University, United States
- Howard Hughes Medical Institute, Northwestern University, United States
- University of Wisconsin, United States
- University of Tübingen, Germany
- Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, United States
Figures
Figure 1 with 2 supplements
CamiCre+; Bmal1fx/fx mice are forebrain/SCN-specific Bmal1 knockouts.
(A) Dorsal view of a CamiCre-; tdTom+ control brain. Bright field. Scale bar: 1 mm. (B) Dorsal view of a CamiCre+; tdTom+ brain. Bright field. (C) Ventral view of (B). (D) Fluorescence image of a …
Figure 1—figure supplement 1
Weights of Bmal1 brain knockout mice are similar to controls at 2 months and 4–6 months of age.
All body weights were measured in live, wheel-naïve animals. Numbers of male mice weighed at 2 months (= 54- to 62-day old) and 4–6 months (= 120- to 195-day old), respectively: Cre (n = 25, 35), …
Figure 1—figure supplement 2
Additional in situ analysis of Bmal1 brain knockout mice.
(A, B) Coronal brain sections at the level of the SCN. Below each coronal brain section is a close-up of the SCN. (A) Mice were sacrificed at CT18 and CT6 following 2 days of DD. Sections were …
Figure 2
Complete loss of circadian rhythmicity of Bmal1 brain knockout mice in constant darkness and constant light.
(A, B) Representative actograms of daily wheel-running activity of Cre, Fx/Fx, Het, and BKO mice. Activity records were double plotted, with each day being represented beneath and also to the right …
Figure 3 with 1 supplement
Both entrainment and masking are abnormal in Bmal1 brain knockout mice.
(A, B) Representative double-plotted actograms of daily wheel-running activity of Cre, Fx/Fx, Het, and BKO mice. Mice were housed in LD, released into DD for 4 weeks, returned to LD for 2 weeks, …
Figure 3—figure supplement 1
Masking in Bmal1 brain knockout mice.
(A) Daytime activity levels during LD, as percentage of total daily activity, in Cre (n = 12), Fx/Fx (n = 18), Het (n = 13), and BKO (n = 16) mice. A significant effect of genotype on daytime …
Figure 4
Real-time reporting of circadian expression of PER2::LUC in the forebrain/SCN knockout mice.
(A) Representative records of bioluminescence reporting of circadian expression from various tissues in Fx/Fx and BKO mice. Tissues were prepared from mice in LD. PMT counts are plotted against the …
Figure 5
Phase analysis of circadian expression of various peripheral tissues from the forebrain/SCN knockout mice.
(A) Phase map of circadian rhythms of various peripheral tissues from Fx/Fx control mice in LD (n = 15, except for pituitary n = 14). The peak phases (or averaged peak phases where more than two …
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Figure 5—source data 1
(A) Summary of statistical analysis of circular plots presented in Figure 5. (B) Statistical analysis of circular variance for peak bioluminescence in individual mice in LD. (C) Statistical analysis of circular variance for peak bioluminescence in individual mice in DD.
- https://doi.org/10.7554/eLife.04617.011
Figure 6 with 1 supplement
Decreased amplitude in peripheral tissues of BKOs.
(A, B) Normalized amplitude of bioluminescence rhythms from Fx/Fx control (open bar) vs BKO (dark bar) in LD (A) and DD (B). Mean relative amplitude ±SD are shown. The sample size is the same as in F…
Figure 6—figure supplement 1
Real-time monitoring of circadian expression in forebrain/SCN knockout mice from DD.
Representative records of bioluminescence reporting of circadian expression from Fx/Fx and BKO mice maintained in DD. Tissues were harvested before the predicted onset of activity (=CT12) of Fx/Fx …
Figure 7 with 1 supplement
Intact FAA in Bmal1 brain knockout mice.
(A) Schedule of a gradual temporal FR protocol in LD (upper) or DD (lower). Horizontal black and white bars represent lights off and on, respectively. Gray bars represent food availability. Since …
Figure 7—figure supplement 1
Lack of Bmal1 expression in the DMH of Bmal1 brain knockout mice.
(A, B) Food consumed during FR was significantly lower in BKOs compared to controls using Repeated Measures GLM ANOVA for both LD conditions [F1,291 = 43.40, p < 0.0001] (A) and DD conditions [F1,293…
Figure 8
Effects of restricted feeding on circadian rhythms of peripheral tissues.
(A) Schedule of a temporal FR protocol in DD for real-time circadian reporting assay. Horizontal black and white bars representing lights off and on for LD 12:12 are shown above as a reference for a …
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Figure 8—source data 1
(A) Summary of statistical analysis of circular plots presented in Figure 8. (B) Statistical analysis of circular variance for peak bioluminescence in individual mice under restriction feeding.
- https://doi.org/10.7554/eLife.04617.017
Figure 9
Analysis of circadian rhythms of peripheral tissues under different conditions.
(A, B) Comparison of peak bioluminescence rhythms in the peripheral tissues in Fx/Fx (A) and BKO (B) mice under LD, DD, and FR conditions. Circular plots are as presented in Figure 5E and Figure 8G. …
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Figure 9—source data 1
(A) Summary of statistical comparison of peak phases from Fx/Fx mice under different conditions. (B) Summary of statistical comparison of peak phases from BKO mice under different conditions.
- https://doi.org/10.7554/eLife.04617.019
