Myofibroblast senescence promotes arrhythmogenic remodeling in the aged infarcted rabbit heart
- Brown University, United States
- Cardiovascular Research Center, Rhode Island Hospital, United States
- Department of Pharmacology, Institute of Graduate Studies in Health Sciences, Istanbul University, Turkey
- School of Medicine, University of California, Los Angeles, United States
- School of Biosciences and Medicine, University of Surrey, United Kingdom
- Department of Hematology, Rhode Island Hospital, United States
- Medical University of South Carolina, United States
Figures
Figure 1
Aged rabbits exhibit increased incidence of peri-procedural arrhythmias.
(A) Survival table of young and aged infarcted rabbits. Procedural deaths were defined as death occurring within the first 3 hr of surgery. (B) Histogram of rabbits included in the study by age at …
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Figure 1—source data 1
Raw data pertaining to rabbits used in the study that was used to create Figure 1.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig1-data1-v3.xlsx
Figure 2
Progression of infarct size, infarct border zone (IBZ) geometry, and fibrosis is consistent between young and aged rabbits.
(A) Left: Whole rabbit heart, with infarct zone outlined. Middle: Dissected left ventricular free wall. Right: Quantification of percent infarcted area of the left ventricular free wall. (B) Left: …
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Figure 2—source data 1
Raw data used to create Figure 2.
Each tab contains raw data for the corresponding panel of Figure 2.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig2-data1-v3.xlsx
Figure 3
The border zone of aged rabbits shows action potential duration (APD) prolongation, APD alternans, and faster ventricular fibrillation (VF) frequency.
(A) Photograph of a rabbit heart showing the infarct zone (IZ), infarct border zone (IBZ), and remote zone (RZ). White line indicates a representative plane along which alternans images in (C) were …
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Figure 3—source data 1
Raw data used to create Figure 3.
Tabs contain raw data corresponding to panel B and panel D.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig3-data1-v3.xlsx
Figure 4 with 1 supplement
Senescence of myofibroblasts is elevated and persistent in the aged rabbit heart post-MI and correlates with increased inflammation.
(A) Representative senescence-associated β-galactosidase (SA-β-gal)-stained image showing examples of infarct zone, border zone, and remote zone (RZ). (B) Representative SA-β-gal-stained images …
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Figure 4—source data 1
Raw data used to create Figure 4.
Tabs contain data corresponding to panel A, panels E and F, panels E and G, and panel H, respectively.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig4-data1-v3.xlsx
Figure 4—figure supplement 1
Senescence assessment of young and aged sham-infarcted rabbits.
(A): Representative senescence-associated β-galactosidase (SA-β-gal)-stained images showing examples of myocardial histology in young and aged rabbits 2 weeks after sham infarction. (B) …
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Figure 4—figure supplement 1—source data 1
Raw data used to create Figure 4—figure supplement 1.
Data corresponds to panel B.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig4-figsupp1-data1-v3.xlsx
Figure 5 with 1 supplement
Presence of senescent myofibroblasts in aged infarcted human ventricular tissue.
(A) Representative images of senescence-associated β-galactosidase (SA-β-gal) (left) and trichrome (right)-stained frozen sections. (B) Representative confocal images of γH2AX/αSMA double …
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Figure 5—source data 1
Raw data used to create Figure 5, panel C.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig5-data1-v3.xlsx
Figure 5—figure supplement 1
Presence of senescent macrophages in aged infarcted human ventricular tissue.
(A) Representative confocal image of γH2AX/CD68 double immunostained frozen section. (B) Inset of (A). Yellow arrows indicate nuclear γH2AX foci.
Figure 6 with 1 supplement
Etoposide induces senescence in adult rabbit cardiac fibroblasts.
(A) Experimental design. (B) Representative senescence-associated β-galactosidase (SA-β-gal)-stained images of treated fibroblasts. (C) Quantification of % SA-β-gal+ cells from (B). (D) …
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Figure 6—source data 1
Raw data used to create Figure 6C.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig6-data1-v3.zip
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Figure 6—source data 2
Raw data used to create Figure 6F.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig6-data2-v3.zip
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Figure 6—source data 3
Raw data used to create Figure 6G.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig6-data3-v3.xls
Figure 6—figure supplement 1
Verification of adult rabbit cardiac fibroblast identity.
Representative images from immunofluorescent staining of adult rabbit cardiac fibroblasts (top) and human aortic endothelial cells (bottom) with myofibroblast and endothelial cell markers.
Figure 7 with 1 supplement
Conditioned media from senescent adult rabbit cardiac fibroblasts does not affect action potential duration (APD) or Ikr.
(A) Representative current traces showing total current, current after application of 5 μM E4031, and their difference that reveals IKr of primary 3-week-old rabbit myocytes treated with conditioned …
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Figure 7—source data 1
Contains raw data used to create Figure 7B.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig7-data1-v3.xlsx
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Figure 7—source data 2
Raw data used to create Figure 7D.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig7-data2-v3.xlsx
Figure 7—figure supplement 1
Twenty-four to 30 hr co-culturing of ventricular cardiomyocytes with senescent or proliferating fibroblasts does not significantly affect action potential duration.
Plots of APD at 90% repolarization (top row) or 50% repolarization (bottom row) during 0.25 Hz pacing (left column) and 1.25 Hz pacing (right column). Quantification was performed on primary …
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Figure 7—figure supplement 1—source data 1
Raw data used to create Figure 7—figure supplement 1.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig7-figsupp1-data1-v3.xlsx
Figure 8 with 1 supplement
Pro-arrhythmic effect of myocytes coupled to senescent fibroblasts compared to non-senescent fibroblasts.
(A) Quantification of the percent of senescence-associated β-galactosidase (SA-β-gal+) cells adjacent to myocytes (left) and representative SA-β-gal-stained images of the infarct border zone (IBZ) …
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Figure 8—source data 1
Raw data used to create Figure 8.
Tabs contain data used to create panels A and B, and panel E, respectively.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig8-data1-v3.xlsx
Figure 8—figure supplement 1
Additional representative images of potential cardiomyocyte-myofibroblast couplings mediated by Cx43.
(A) Image of another aged rabbit infarct border zone (IBZ) 3 weeks post-MI, with staining and imaging parameters identical to those presented in Figure 8D. (B) Inset of (A). (C) and (D) Additional …
Figure 9 with 1 supplement
Computational modeling of myocyte-myofibroblast interactions.
(A) Quantification of proliferating and senescent myofibroblast volume from immunofluorescent staining against WGA and γH2AX of frozen tissue sections (left) and adult rabbit cardiac fibroblasts in …
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Figure 9—source data 1
Raw data used to create Figure 9.
- https://cdn.elifesciences.org/articles/84088/elife-84088-fig9-data1-v3.xlsx
Figure 9—figure supplement 1
Additional parameters for myocyte-myofibroblast modeling.
(A) Representative current traces of proliferating and senescent adult rabbit cardiac myofibroblasts at various membrane potentials. (B) I-V curves of cumulative data from proliferating and …
Additional files
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Supplementary file 1
Antibodies used for immunofluorescence staining.
- https://cdn.elifesciences.org/articles/84088/elife-84088-supp1-v3.xlsx
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Supplementary file 2
Rabbit-specific primers used for qPCR.
- https://cdn.elifesciences.org/articles/84088/elife-84088-supp2-v3.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/84088/elife-84088-mdarchecklist1-v3.docx
