Terminal Warm Blood Cardioplegia Improves Cardiac Function Through Microtubule Repolymerization Hironori Tenpaku, MD, Koji Onoda, MD, Kyoko Imanaka-Yoshida,

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Presentation transcript:

Terminal Warm Blood Cardioplegia Improves Cardiac Function Through Microtubule Repolymerization Hironori Tenpaku, MD, Koji Onoda, MD, Kyoko Imanaka-Yoshida, MD, Toshimichi Yoshida, MD, Takatsugu Shimono, MD, Hideto Shimpo, MD, Isao Yada, MD The Annals of Thoracic Surgery Volume 65, Issue 6, Pages 1580-1587 (June 1998) DOI: 10.1016/S0003-4975(98)00237-9

Fig 1 Changes in cardiac function in isolated perfused rat hearts. (a) Changes in heart rate; (b) Changes in coronary flow; (c) Changes in left ventricular developed pressure. Values are expressed as the percent recovery compared with control values and represent the mean ± standard error of the mean for six hearts. (∗p < 0.01 compared with group C at the same time point.) The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)

Fig 2 Immunohistochemical staining of microtubules from isolated perfused rat hearts from group C. (a) Longitudinal section from a control rat heart. Microtubules are seen as a filamentous network in the cytoplasm and a circular formation around the nuclei. (b) A section from a heart during hypothermic arrest. Microtubules are not seen in this section. (c) A section from a heart 5 minutes after reperfusion. Microtubules have repolymerized around the nuclei. (d) A section from a heart 10 minutes after reperfusion. The cytoplasmic microtubules forming the reticular filaments has been reconstructed. (e) A section from a heart 60 minutes after reperfusion. The microtubules are similar to those seen in control hearts. Scale bar represents 10 μm. The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)

Fig 3 Double staining for microtubules and actin filaments in isolated perfused rat hearts. (a and b) Longitudinal sections from hearts during hypothermic cardiac arrest. (a) Staining for microtubules. Microtubules are not seen in about half of the area of the tissue sections analyzed (asterisk). (b) Staining for actin filaments by rhodamine-phalloidin localizes to the cross striations of the sarcomeres. No morphologic changes in the actin filaments are seen during hypothermic cardiac arrest. Scale bar represents 10 μm. The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)

Fig 4 Immunohistochemical staining of microtubules from isolated perfused rat hearts from group TC. (a) Longitudinal section from a heart 5 minutes after reperfusion. (b) A section from a heart 10 minutes after reperfusion. (c) A section from a heart 60 minutes after reperfusion. The morphology of the microtubules is similar to that in hearts from group C at the same time point. Scale bar represents 10 μm. The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)

Fig 5 Densitometric quantification of immunoblots for free and polymerized tubulin fractions from isolated rat left ventricles before and after hypothermic arrest and after reperfusion. (a) Changes in free tubulin. (b) Changes in polymerized tubulin. Values are reported as the mean ± standard error of the mean for four experiments. (∗p < 0.05 versus the respective control value; †p < 0.05 versus the respective control value.) The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)

Fig 6 Effect of colchicine on cardiac function in isolated perfused rat hearts. (a) Changes in heart rate; (b) Changes in coronary flow; (c) Changes in left ventricular developed pressure. Values are expressed as the percentage compared with control values and represent the mean ± standard error of the mean for four hearts. (∗p < 0.01 compared with group N at the same time point.) The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)

Fig 7 Immunoblot analysis of free and polymerized tubulin fractions and immunohistochemical staining of microtubules from isolated perfused rat hearts 60 minutes after perfusion with Krebs-Henseleit buffer or with Krebs-Henseleit buffer containing colchicine. (a) Immunoblots of free tubulin fraction. (b) Immunoblots of polymerized tubulin fraction. Lanes 1 and 6, purified bovine brain tubulin; lanes 2 and 4, tubulin from group N; lanes 3 and 5, tubulin from group CO. (c) Longitudinal section from a heart in group N. The morphology of the microtubules is similar to that in control hearts (see Fig 2a). (d) Longitudinal section from a heart in group CO. Cytoplasmic microtubules have decreased; however, microtubules around the nuclei have not disappeared. Arrows indicate nuclei. Scale bar represents 10 μm. The Annals of Thoracic Surgery 1998 65, 1580-1587DOI: (10.1016/S0003-4975(98)00237-9)