Myocyte apoptosis occurs early during the development of pressure-overload hypertrophy in infant myocardium Yeong-Hoon Choi, MD, Douglas B. Cowan, PhD,

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Myocyte apoptosis occurs early during the development of pressure-overload hypertrophy in infant myocardium Yeong-Hoon Choi, MD, Douglas B. Cowan, PhD, Adrian M. Moran, MBBS, Steven D. Colan, MD, Christof Stamm, MD, Koh Takeuchi, MD, Ingeborg Friehs, MD, Pedro J. del Nido, MD, Francis X. McGowan, MD The Journal of Thoracic and Cardiovascular Surgery Volume 137, Issue 6, Pages 1356-1362.e3 (June 2009) DOI: 10.1016/j.jtcvs.2008.12.020 Copyright © 2009 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Effects of aortic banding of infant rabbits on LV mass:volume ratio determined by serial echocardiography. Data are mean ± standard deviation (SD), n = 6–10 each. #P < .05 versus age-matched controls. ∗P < .05 versus 4-week-old banded. LV mass:volume ratio increases early in hypertrophy and then progressively declines. LV, Left ventricular. The Journal of Thoracic and Cardiovascular Surgery 2009 137, 1356-1362.e3DOI: (10.1016/j.jtcvs.2008.12.020) Copyright © 2009 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Effects of aortic banding of infant rabbits on LV peak systolic stress. Data are mean ± SD, n = 6–10 each. Peak systolic stress is reduced in banded animals at 4 weeks in association with increased LV mass (see Figure 1). Failure of continued hypertrophy in the face of increasing afterload results in a progressive increase in peak systolic stress. ∗P < .05 versus controls. PkSS, Peak systolic stress. The Journal of Thoracic and Cardiovascular Surgery 2009 137, 1356-1362.e3DOI: (10.1016/j.jtcvs.2008.12.020) Copyright © 2009 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Apoptosis in LV myocardium of control sham-operated and banded infant rabbits. A, Representative fluorescent photomicrographs of 5-μm sections of the LV wall from banded rabbits. Sections were labeled with Hoechst to stain nuclear DNA (blue), desmin to identify cardiomyocytes (red), and DNA strand breaks (TUNEL-green). After image acquisition, images were subjected to 2-dimensional “no neighbors” deconvolution using a kernel that estimates the 3-dimensional point spread function based on the optics and imaging system. A and B, Banded animal at 3 weeks. C and D, Banded animal at 7 weeks. Apoptotic nuclei localized to myocytes (arrows); TUNEL-positive staining in non-myocytes (asterisks). B, Assessment of myocyte apoptosis in sham versus banded animals using this approach. Data are mean ± SD, n = 6–8 each. The difference between sham-operated and banded animals was significant at all ages. The Journal of Thoracic and Cardiovascular Surgery 2009 137, 1356-1362.e3DOI: (10.1016/j.jtcvs.2008.12.020) Copyright © 2009 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 LV myocardial caspase-3 activity measured by cleavage of the specific fluorescent substrate DEVD-AFC (BioMol, Plymouth Meeting, Pa) in age-matched sham operated control (open bars) and banded hypertrophying (solid bars) hearts. Data are mean ± SD, n = 5–7 per group and expressed as arbitrary fluorescent units relative to that measured in 7-week controls. ∗P = .04. #P = .03. +P = .008. The Journal of Thoracic and Cardiovascular Surgery 2009 137, 1356-1362.e3DOI: (10.1016/j.jtcvs.2008.12.020) Copyright © 2009 The American Association for Thoracic Surgery Terms and Conditions