Erin E. Tansey, BS, Kevin F. Kwaku, MD, PhD, Peter E

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Presentation transcript:

Reduction and Redistribution of Gap and Adherens Junction Proteins After Ischemia and Reperfusion Erin E. Tansey, BS, Kevin F. Kwaku, MD, PhD, Peter E. Hammer, MS, Douglas B. Cowan, PhD, Micheline Federman, PhD, Sidney Levitsky, MD, James D. McCully, PhD The Annals of Thoracic Surgery Volume 82, Issue 4, Pages 1472-1479 (October 2006) DOI: 10.1016/j.athoracsur.2006.04.061 Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 (A) Histochemical analysis. Representative Masson Trichrome (Panel A) and hematoxylin and eosin (Panel B) stained myocardial sections (×100 original magnification) of left ventricular tissue sections from control and hearts after 30 minutes of global ischemia (GI-30). Scale bars are shown in microns. Histochemistry shows control hearts with preserved myofilament structure and GI-30 hearts with longitudinal and transverse interfibrillar separation. (B) Interfibrillar space. Quantitative light microscopy spatial analysis was performed at low magnification (×100 original magnification) covering the full microscopic slide specimen area using Metamorph Imaging Analysis software and expressed as interfibrillar space in arbitrary units (AU). All results are shown as mean ± SEM for n = 30 for each sample. Significant differences are shown as *p < 0.001 versus control; **p < 0.001 versus GI-20; and ***p < 0.001 versus GI-25. The Annals of Thoracic Surgery 2006 82, 1472-1479DOI: (10.1016/j.athoracsur.2006.04.061) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Immunohistochemical analysis. Representative confocal photomicrographs (×40 original magnification) of immunohistochemically stained left ventricular free wall sections from control and hearts after 15 and 30 minutes of global ischemia (GI). Overlay shows nuclei as blue, myocytes as red (F-actin stained with Alexa568-phalloidin), and N-cadherin (A), integrin-β1, (B) and Cx43[α1] (C) as green. Scale bars are shown in microns. Photomicrographs show a qualitative decrease in N-cadherin and Cx43[α1] at GI-15 and GI-30. Cx43[α1] was localized predominantly at the intercalated disc in control, but was redistributed to the periphery of the myocyte after more than 15 minutes global ischemia. There was no change in integrin-β1. The Annals of Thoracic Surgery 2006 82, 1472-1479DOI: (10.1016/j.athoracsur.2006.04.061) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Western blot analysis. Representative Western blots for N-cadherin, integrin-β1, and Cx43[α1] and Cx43[α1] phosphorylated Cx43[α1] (46 kDa) and dephosphorylated Cx43[α1] (41 kDa). Molecular weights (kDa) for each band are shown. Densitometric analysis for n = 4 blots each is shown on the right. Results show a significant decrease in N-cadherin at 15 minutes of global ischemia (GI) and Cx43[α1] and phosphorylated Cx43[α1] (46kDa) at GI-20 (p < 0.0001 versus control). There were no significant changes in integrin-β1 All results are shown as mean ± SEM. Significant differences are shown as *p < 0.001 versus control, and **p < 0.001 versus GI-20. The Annals of Thoracic Surgery 2006 82, 1472-1479DOI: (10.1016/j.athoracsur.2006.04.061) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Conduction velocity (CV) analysis. Epicardial conduction velocities were measured at 30 (equilibrium), 60, 70, 90, 120, and 180 minutes of perfusion along the longitudinal (CVL) and transverse (CVT) axes of epicardial myocardial fiber orientation. Anisotropic ratios (AR) were calculated as CVL/CVT. No significant change was seen in CVL, CVT or AR between global ischemia (GI) after 30 minutes and control groups at equilibrium or for the duration of perfusion time (p = 0.903 for CVL; p = 1.0 for CVL; p = 0.586 for AR). All results are shown as mean ± SEM for n = 5 in each group. The Annals of Thoracic Surgery 2006 82, 1472-1479DOI: (10.1016/j.athoracsur.2006.04.061) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions