Loss of the melanocortin-4 receptor in mice causes dilated cardiomyopathy
- Vanderbilt University, United States
- Department of Medicine, Vanderbilt University, United States
- University of Michigan, United States
- Vanderbilt University Department of Medicine, United States
- Vanderbilt University Medical Center, United States
- University of Michigan School of Medicine, United States
Figures
Figure 1
Mc4r deletion causes progressive cardiac dilatation and reduced contractility.
(A) Representative B-mode Parasternal long axis (PSLAX) image at diastole (top) and M-mode short axis (SAX) image of male WT C57B6/J mouse at 26 weeks of age. (B) Representative B-mode PSLAX image …
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Figure 1—source data 1
Source data for Figure 1C–G and J-N.
This source file provides mean, SD, sample size and statistical test information for all echocardiography analyses in Figure 1, of which examples are shown in Figure 1A–B and H-I.
- https://doi.org/10.7554/eLife.28118.004
Figure 2
Mc4r−/− animals display bradycardic arrhythmias.
Rhythm strips of 30 week old WT, Mc4r-/+ and Mc4r−/− animals taken during echocardiograms. Mc4r−/− mice display sinus bradycardia with abnormal rhythmicity.
Figure 3
Mc4r loxTB animals phenocopy the reduced cardiac function seen in Mc4r−/− mice.
(A) Representative B-mode PSLAX image at diastole (top) and M-mode SAX image of male Mc4r loxTB -/- (WT) mouse at 35 weeks of age. (B) Representative B- mode PSLAX image at diastole (top) and M-mode …
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Figure 3—source data 1
Source data for Figure 3C–G.
This source file provides mean, SD, sample size and statistical test information for all echocardiography analyses in Figure 3, of which examples are shown in Figure 3A–B.
- https://doi.org/10.7554/eLife.28118.007
Figure 4
Mc4r is not expressed in adult heart tissue.
(A) qRT-PCR comparison of Mc4r expression in heart and hypothalamus using the ΔΔCT method. (Student’s t-test, p<0.0001, n = 8) (B) qRT-PCR comparison of Mc4r expression in the hypothalamus and …
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Figure 4—source data 1
Source data for Figure 4A–B.
This source file provides mean, SD, sample size and statistical test information for all qRT-PCR experiments in Figure 4.
- https://doi.org/10.7554/eLife.28118.009
Figure 5
Age and weight matched wild type animals fed a high fat diet do not phenocopy the cardiomyopathy seen in Mc4r−/− mice.
(A) Body weights of age matched WT, diet induced obese (DIO), and Mc4r−/− mice. (1-way ANOVA Sidak multiple comparison test, p<0.0001, n = 5). (B) Lean mass of WT, DIO and Mc4r−/− mice by NMR (1-way …
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Figure 5—source data 1
Source data for Figure 5A–G and K-O.
This source file provides mean, SD, sample size and statistical test information for all body compostion, glucose tolerance, and echocardiography studies in Figure 5.
- https://doi.org/10.7554/eLife.28118.011
Figure 6
Histological analysis of Mc4r−/− hearts reveals mitochondrial pleomorphy and cardiomyocyte dropout.
(A) Representative gross pathology of WT and Mc4r−/− hearts (B) Heart wet weight of 6, 12 and 30 week old WT and Mc4r−/− animals (Student’s t-test, p<0.01, n = 4–7) (C) Heart weight to total lean …
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Figure 6—source data 1
Source data for Figure 6B–D and 6F.
This source file provides mean, SD, sample size and statistical test information for the gross pathological and histological analyses presented in Figure 6.
- https://doi.org/10.7554/eLife.28118.013
Figure 7
Mc4r−/− heart tissue displays increased mitochondrial capacity.
(A) Relative qPCR of mitochondrial DNA content using H19 and CytB primers (Students t-test of ΔCT values, p>0.05, n = 6). (B) Representative western blot of whole cell tissue lysates from WT and Mc4r…
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Figure 7—source data 1
Source data for Figure 7A and C, and 7E-H.
This source file provides mean, SD, sample size and statistical test information for all studies presented in Figure 7.
- https://doi.org/10.7554/eLife.28118.015
Figure 8
Mitochondrial capacity changes in Mc4r−/− animals are present in young animals and specific to the heart.
(A) O2 consumption rate comparison between WT and Mc4r−/− myocardial tissue from 6 to 12 week old mice. (2 way ANOVA Sidak post test, p<0.05, n = 7) at 6–12 weeks of age (B) ADP dependent O2 …
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Figure 8—source data 1
Source data for Figure 8A–F.
This source file provides mean, SD, sample size and statistical test information for all high resolution respirometry studies in Figure 8.
- https://doi.org/10.7554/eLife.28118.017
Figure 9
Red Gastrocnemius Ox-Phos components.
(A) Relative quantification of ETC blot normalized to ATP synthase content in WT tissue (2 way ANOVA Bonferroni post test, p>0.05, n = 3 blots x 5 samples per genotype) (B) Representative western …
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Figure 9—source data 1
Source data for Figure 9A.
- https://doi.org/10.7554/eLife.28118.019
Figure 10
Mc4r−/− hearts display signs of increased ROS production.
(A) DCF assay comparing myocardial lysates from 6 to 12 week old Mc4r−/− and WT mice (ratio paired t-test, p<0.05, n = 5). (B) DCF assay comparing myocardial lysates from 30 to 33 week old Mc4r−/− …
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Figure 10—source data 1
Source data for Figure 10A–D.
This source file provides mean, SD, sample size and statistical test information for all ROS studies presented in Figure 10.
- https://doi.org/10.7554/eLife.28118.021
Figure 11
Gene changes in Mc4r−/− myocardium.
(A) Volcano plot of RNA-seq results. Red points indicate gene changes that reached statistical significance (n = 4 WT, n = 6 Mc4r−/−). (B) qRT-PCR confirmation of select gene changes found with …
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Figure 11—source data 1
Source data for Figure 11B.
This source file provides mean, SD, sample size and statistical test information for the qRT-PCR studies presented in Figure 11B.
- https://doi.org/10.7554/eLife.28118.023
Figure 12
Mc4r ± mice are hypersensitive to doxorubicin.
(A) Treatment strategy to subclinical cardiac dysfunction in WT C57B6/J mice. Mice were given 5 mg/kg doxorubicin over the course of 2 weeks during which time their bodyweights were recorded. (B) …
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Figure 12—source data 1
Source data for Figure 12B.
This source file provides mean, SD, sample size and statistical test information for the doxorubicin studies outlined in Figure 12A and presented in Figure 12B.
- https://doi.org/10.7554/eLife.28118.025
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.28118.026
