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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Circulation Research. 2010;106:354-362, originally published February 4, 2010
https://doi.org/10.1161/CIRCRESAHA.109.207423

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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

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Table 1.
By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Non-standard Abbreviations and AcronymsShow More

Non-standard Abbreviations and AcronymsShow Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 1. PLN ablation in CaMKIIδc TG mice repletes SR Ca2+ load in isolated cardiomyocytes. A, Representative caffeine-induced Ca2+ transient traces....Show More

Figure 1. PLN ablation in CaMKIIδc TG mice repletes SR Ca2+ load in isolated cardiomyocytes. A, Representative caffeine-induced Ca2+ transient traces. B, Average Δ[Ca2+]i for caffeine-induced Ca2+ transients (SR Ca2+ load). *P<0.05 vs WT. #P<0.05 vs CaMKII-TG. C, Average rate constant kCa for [Ca2+]i decline during caffeine exposure (which indicates NCX activity). *P<0.05 vs WT; **P<0.01 vs WT. The number of cells studied: WT n=6, PLN-KO n=21, CaMKII-TG n=9, and KO/TG n=9.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 2. PLN ablation in CaMKIIδc TG mice normalizes twitch Ca2+ transients in isolated cardiomyocytes. A, Representative twitch [Ca2+]i transient tr...Show More

Figure 2. PLN ablation in CaMKIIδc TG mice normalizes twitch Ca2+ transients in isolated cardiomyocytes. A, Representative twitch [Ca2+]i transient traces. B, Average Δ[Ca2+]i for twitch-induced Ca2+ transients. *P<0.05 vs WT; #P<0.05 vs CaMKII-TG. The number of cells studied: WT n=6, PLN-KO n=21, CaMKII-TG n=9, and KO/TG n=9.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 3. PLN ablation in CaMKIIδc TG mice exaggerates the phenotype of dilated cardiomyopathy. A, Heart weight/body weight (HW/BW) ratio at 8 weeks....Show More

Figure 3. PLN ablation in CaMKIIδc TG mice exaggerates the phenotype of dilated cardiomyopathy. A, Heart weight/body weight (HW/BW) ratio at 8 weeks. B and C, Averaged echocardiographic parameters at different ages. Data are presented for B, left ventricular end-diastolic diameter (LVEDD), and for C, calculated percent fractional shortening (FS). n=4 to 6 mice per group at each age; *P<0.05 vs WT. #P<0.05 vs CaMKII-TG. D, Survival curves for CaMKII-TG and PLN-KO/CaMKII-TG mice. PLN ablation accentuates premature death in CaMKIIδC TG mice. Numbers of mice were as follows: CaMKII-TG, n=16; KO/TG, n=22. P<0.01 KO/TG vs CaMKII-TG.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 4. Quantitative immunoblotting of major Ca2+ handling proteins in mouse ventricular homogenates. A, Expression levels of calsequestrin (CsQ), S...Show More

Figure 4. Quantitative immunoblotting of major Ca2+ handling proteins in mouse ventricular homogenates. A, Expression levels of calsequestrin (CsQ), SERCA, and NCX did not change significantly between the CaMKII-TG and the PLN-KO/CaMKII-TG. B, Phosphorylation of ryanodine receptors (RyR2) at Ser2809 and Ser2815 (normalized to total RyR2) was increased significantly but not differentially in CaMKIIδc TG and PLN-KO/CaMKII-TG mice. *P<0.05 vs WT; n=4 mice per group.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 5. Enhanced SR Ca2+ spark frequency and Ca2+ leak in intact cardiomyocytes from PLN-KO/CaMKII-TG mice. A, SR Ca spark frequency (CaSpF). *P<...Show More

Figure 5. Enhanced SR Ca2+ spark frequency and Ca2+ leak in intact cardiomyocytes from PLN-KO/CaMKII-TG mice. A, SR Ca spark frequency (CaSpF). *P<0.05 vs WT; #P<0.05 vs CaMKII-TG; +P<0.05 vs KO/TG. B, SR Ca2+ leak. *P<0.05 vs WT; #P<0.05 vs CaMKII-TG; +P<0.05 vs KO/TG.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 6. Apoptosis is greater in PLN-KO/CaMKII-TG than in CaMKIIδc TG cardiomyocytes. A, Representative TUNEL staining images in CaMKII-TG and PLN-KO...Show More

Figure 6. Apoptosis is greater in PLN-KO/CaMKII-TG than in CaMKIIδc TG cardiomyocytes. A, Representative TUNEL staining images in CaMKII-TG and PLN-KO/CaMKII-TG mouse heart sections. B, Average TUNEL positive myocytes. *P<0.05 vs WT; n=4 mice per group. Data were normalized to WT; the average number from all experiments was ≈75 TUNEL-positive nuclei/105 myocytes in the KO/TG group.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 7. Inhibition of SR Ca2+ leak by KN-93 or ryanodine decreases the rate of death in isolated cardiomyocytes from PLN-KO/CaMKII-TG mice. Isolated...Show More

Figure 7. Inhibition of SR Ca2+ leak by KN-93 or ryanodine decreases the rate of death in isolated cardiomyocytes from PLN-KO/CaMKII-TG mice. Isolated cardiomyocytes were plated and incubated in the presence and absence of inhibitors, and cell viability was assessed at various times using morphology and at 12 hours by trypan blue exclusion. A, WT myocytes showed only slight decreases in viability over 12 hours after plating. B and C, The percent of living myocytes from both CaMKII-TG and PLN-KO/CaMKII-TG mice decreased dramatically over 12 hours. Inhibition of CaMKII with KN-93 or of SR Ca2+ leak with ryanodine decreased the rate of myocyte death in both CaMKII-TG and PLN-KO/CaMKII-TG mice.Show Less
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Phospholamban Ablation Rescues Sarcoplasmic Reticulum Ca²⁺ Handling but Exacerbates Cardiac Dysfunction in CaMKIIδ_C Transgenic Mice
Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers, Joan Heller Brown

Circulation Research February 2010, 106 (2) 354-362; DOI: https://doi.org/10.1161/CIRCRESAHA.109.207423

By Tong Zhang, Tao Guo, Shikha Mishra, Nancy D. Dalton, Evangelia G. Kranias, Kirk L. Peterson, Donald M. Bers and Joan Heller Brown

Figure 8. Involvement of mitochondrial Ca2+ in myocyte death in PLN-KO/CaMKII-TG mice. A, Mitochondrial Ca2+ was measured as shown in the protocol. NT...Show More

Figure 8. Involvement of mitochondrial Ca2+ in myocyte death in PLN-KO/CaMKII-TG mice. A, Mitochondrial Ca2+ was measured as shown in the protocol. NT indicates no treatment; Thaps, thapsigargin. B, Mitochondrial Ca2+ was increased in cardiomyocytes isolated from PLN-KO/CaMKII-TG mice. *P<0.05 vs WT. Mitochondrial Ca2+ in PLN-KO/CaMKII-TG myocytes returned to control levels with reduced external Ca2+ (0.3 mmol/L) and CaMKII inhibitor AIP. C, Reduced external Ca2+ and CaMKII inhibition with AIP returned SR Ca2+ content and Ca2+ transients to near control myocyte levels. D, Inhibition of the mitochondrial uniporter by Ru-360 or inhibition of the mitochondrial permeability transition pore by cyclosporin A decreased the rate of death in isolated cardiomyocytes from PLN-KO/CaMKII-TG mice at 12 hours. *P<0.05 vs WT.Show Less