Genetic deletion of calcium/calmodulin-dependent protein kinase type II delta does not mitigate adverse myocardial remodeling in volume-overloaded hearts

Figures

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  Figure 1. CaMKIIδ deletion does not improve mortality rates post-shunt. (a–c) Comparable left ventricle dilatation, stroke volume and cardiac output at 1-week post-shunt in both genotypes. (d) Kaplan-Meier survival curves depicting the effect of CaMKIIδ deletion on 10-week overall survival in VO. *p < 0.05, **p < 0.01, ***p < 0.001 vs. corresponding sham, 1-way ANOVA with Bonferroni post-test (a-c) and log-rank test (d). Numbers within columns indicate mice.

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  Figure 2. Altered myocardial geometry and systolic impairment are not different in δ-KO and WT hearts at 12 weeks of VO. (a) Representative M-mode echocardiogram. (b–g) Parameters of cardiac structure and function. Data are presented as mean ± SEM. *p < 0.05, ***p < 0.001 vs. corresponding sham, 1-way ANOVA with Bonferroni post-test. Numbers within columns indicate mice. bpm, beats/minute; CO, cardiac output; EF, ejection fraction; HR, heart rate; LVEDD, left ventricular end-diastolic diameter; LVM/EDV, left ventricular mass-to-end-diastolic volume.

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  Figure 3. Speckle tracking echocardiography reveals systolic and diastolic dysfunction in δ-KO and WT littermates after shunt. (a) Mean global longitudinal strain rate. (b) Radial diastolic peak velocity. (c) Average reverse longitudinal strain rate (rLSR). Data are expressed as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 vs. corresponding sham, 1-way ANOVA with Bonferroni post-test. Numbers within columns indicate mice.

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  Figure 4. Cardiomyocytes hypertrophy and induced apoptosis occurs in δ-KO and WT hearts at 12 weeks of VO. (a) Representative cardiac transverse sections stained with hematoxylin and eosin (H&E). (b) Picrosirius red stained sections are illustrated (left panels) and quantification of fibrosis (right panel). (c) Stained cardiac sections with wheat germ agglutinin (WGA, left panels) and quantification of myocytes minimal fiber diameter (right panel). (d) Representative photomicrographs showing TUNEL staining (left panels) and quantification of TUNEL-positive cells (right panel). Positive (DNase I treatment) and negative (without TUNEL enzyme) controls were also shown. Data are presented as mean ± SEM. *p < 0.05 vs. corresponding sham, 1-way ANOVA with Bonferroni post-test. Numbers within columns indicate mice.

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  Figure 5. CaMKIIδ deletion does not alter the expression of cardiac stress markers at 12 weeks post-shunt. Quantitative real-time PCR analyses of fetal cardiac genes expression. Data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 vs. corresponding sham, 1-way ANOVA with Bonferroni post-test. Numbers within columns indicate mice. Nppa, natriuretic peptide type A; Nppb, natriuretic peptide type B; Serca2a, sarcoplasmic reticulum Ca2+-ATPase-2a.

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  Figure 6. Analysis of CaMKII targeted proteins in δ-KO and WT hearts at 12 weeks of VO. (a) Representative Immunoblots and Ponceau-stained blots from left ventricles of δ-KO and WT hearts (Uncropped blots are shown in Fig. S1). (b) Bands intensity was normalized to Ponceau staining. Analysis per heart was performed in duplicates. (c) mRNA expression of CamkII isoforms. Data are mean ± SEM. *p < 0.05 vs. corresponding sham, 1-way ANOVA with Bonferroni post-test. Numbers within columns indicate mice.