Shigeto Kanno

The role of myocardial gap junctions in electrical conduction and arrhythmogenesis

Electrical activation of the heart requires cell-cell transfer of current via gap junctions, arrays of densely packed protein channels that permit intercellular passage of ions and small molecules. Because current transfer occurs only at gap junctions, the spatial distribution and biophysical properties of gap junction channels are important determinants of the conduction properties of cardiac muscle. Gap junction channels are composed of members of a multigene family of proteins called connexins. As a general rule, individual cells express multiple connexins, which creates the potential for considerable functional diversity in gap junction channels. Although gap junction channels are relatively nonselective in their permeability to ions and small molecules, cardiac myocytes actively adjust their level of coupling by multiple mechanisms including changes in connexin expression, regulation of connexin trafficking and turnover, and modulation of channel properties. In advanced stages of heart disease, connexin expression and intercellular coupling are diminished, and gap junction channels become redistributed. These changes have been strongly implicated in the pathogenesis of lethal ventricular arrhythmias. Ongoing studies in genetically engineered mice are revealing insights into the role of individual gap junction channel proteins in normal cardiac function and arrhythmogenesis.

Connexin43 as a determinant of myocardial infarct size following coronary occlusion in mice

OBJECTIVES The purpose of this study was to define the role of cell-cell coupling as an independent determinant of infarct size following coronary occlusion. BACKGROUND Electrical uncoupling induced by acute ischemia enhances arrhythmogenesis, but it may also protect the heart by limiting intercellular spread of chemical mediators of injury. METHODS The left anterior descending coronary artery was ligated in wild-type (Cx43(+/+)) mice and Cx43-deficient (Cx43(+/-)) mice that are heterozygous for a null allele in the gene encoding the major gap junction channel protein, connexin43 (Cx43). Ventricular remodeling and infarct size were compared in both groups. RESULTS Echocardiography at 1 and 10 weeks after infarction showed that left ventricular end-diastolic volume and mass increased and ejection fraction decreased in proportion to infarct size in both Cx43(+/-) and Cx43(+/+) hearts. However, infarct size measured histologically in healing infarcts (eight days after infarction) was 29% smaller in Cx43(+/-) hearts (17 +/- 14% of total left ventricular area, n = 30) than in Cx43(+/+) hearts (24 +/- 15%, n = 23; p = 0.037). Fully healed infarcts were smaller than healing infarcts, owing to resorption of necrotic tissue and maturation of scar, but infarct size at 10 weeks after coronary occlusion was still smaller (by 50%) in Cx43(+/-) hearts (6 +/- 5%, n = 9) compared with Cx43(+/+) hearts (12 +/- 7%, n = 17; p = 0.037). CONCLUSIONS Cx43-deficient mice develop smaller infarcts than wild-type mice following coronary ligation. New therapies designed to decrease the risk of arrhythmias by enhancing intercellular communication could lead to larger infarcts caused by persistent coronary occlusion.

Remodeling of Gap Junctions in Ischemic and Nonischemic Forms of Heart Disease

Electrical activation of the myocardium to produce effective pumping of blood depends on the orderly coordinated spatial and temporal transfer of current from one cell to another via gap junctions. Normal ventricular myocytes are extensively coupled by gap junctions and have the capacity to rapidly increase the amount of connexin within gap junction plaques to meet physiological demands for enhanced cell-cell communication. However, myocytes can also rapidly uncouple in response to injury or disease. In general, both acute and chronic forms of heart disease caused by diverse etiologies are associated with changes in the expression of connexins and remodeling of gap junctions. Such remodeling may have both adaptive and maladaptive consequences and contribute to major clinical processes such as heart failure and sudden cardiac death. Our laboratory has investigated mechanisms regulating cell-cell electrical coupling in the heart under physiological and pathophysiological conditions. This review is focused on selected aspects of this work pertaining to changes in coupling in response to acute and chronic ischemic heart disease and in familial cardiomyopathies caused by mutations in genes encoding desmosomal proteins.

Stromal cell‐derived factor‐1α improves infarcted heart function through angiogenesis in mice

Background: Local delivery of stromal cell‐derived factor‐1α (SDF‐1) has been demonstrated to improve hind limb ischemia through enhanced neovascularization in animals. It was hypothesized that local administration of SDF‐1 also contributes to neovascularization of ischemic heart.

Spontaneous and inducible ventricular arrhythmias after myocardial infarction in mice.

INTRODUCTION Remodeling of gap junctions has been implicated in development of ventricular arrhythmias following myocardial infarction (MI) but the specific contribution of reduced electrical coupling is not known. We addressed this question using hearts from mice heterozygous for a connexin43 null allele (Cx43(+/-)). METHODS To determine whether Cx43-deficient mice exhibit increased spontaneous ventricular arrhythmias in the setting of chronic ischemic heart disease, radiofrequency transmitters were implanted in wild-type and Cx43(+/-) mice 2 days or 9 weeks after left anterior descending coronary artery ligation or sham operations. ECGs were recorded from unanesthetized, unrestrained mice 1 and 10 weeks after MI. Isolated, perfused hearts excised 1 and 10 weeks after MI were subjected to programmed electrical stimulation to induce arrhythmias. RESULTS AND CONCLUSIONS Hearts with infarcts exhibited more spontaneous and inducible arrhythmias, but there was no significant difference between wild-type and Cx43-deficient mice. Fewer hearts exhibited spontaneous ventricular tachycardia (VT) in vivo than were inducible in vitro, suggesting that structural and functional substrates for inducible VT in isolated hearts may not be sufficient for initiation and maintenance of sustained VT in vivo. Previous studies have shown that Cx43-deficient mice exhibit more VT than wild-type mice during acute regional ischemia. Mice with MI exhibit increased arrhythmias. However, reduced coupling in Cx43-deficient mice does not significantly enhance spontaneous or inducible VT after MI.

Fibronectin and tenascin in rat tracheal wound healing and their relation to cell proliferation.

To investigate the relationship between cell proliferation and distribution of fibronectin and tenascin during wound healing, light and electron microscopy and immunohistochemistry for fibronectin, tenascin, and 5‐bromodeoxyuridine (BrdU) were performed following mechanical injury of rat trachea. Tenascin staining appeared 18h after curettage, when the percentage of BrdU‐positive nuclei was maximal in the epithelium. Once tenascin appeared, the labeling index of BrdU‐positive epithelial nuclei decreased rapidly. Distribution of tenascin was restricted to granulation tissue In curetted areas which were covered with regenerating epithelium, while fibronectin stained diffusely in both curetted and non‐curetted areas. Analysis of the relative intensity of fibronectin and tenascin staining showed that decreases of fibronectin staining were followed by increasing tenascin staining. It is proposed that fibronectin and tenascin may contribute differently to tissue repair in the trachea by interfering with cell proliferation of epithelial cells and fibroblasts.

Impact of sequential grafting of the internal thoracic or right gastroepiploic arteries on multiple coronary revascularization.

OBJECTIVE The aim of the study is to clarify the efficacy of the sequential anastomotic technique of the arterial conduits for multiple coronary revascularization. BACKGROUND The internal thoracic artery (ITA) is now widely accepted as a durable conduit for myocardial revascularization. The right gastroepiploic artery (GEA) has been developed as a third in situ arterial graft with an outcome similar to that of the ITA. MATERIAL AND METHOD One hundred and forty five consecutive patients (116 male, 29 female, mean age 60.4yr) who received sequential grafting of either the ITA or GEA or both were retrospectively analysed. RESULTS Sequential anastomoses were performed in 121 in situ left ITAs, 36 in situ GEAs and 12 composite right ITAs. No in situ right ITA was anastomosed sequentially. Two to six vessels (mean 3.8) were revascularized for each patient. Of the total 543 bypassed vessels, 432 (79.6%) were reconstructed with the arterial grafts. In 85 patients with quadruple bypass or more, the arterial grafts were able to reconstruct 266 out of 360 (74.0%) target vessels. Seventy one patients (49.0%) were revascularized without venous grafts. The arterial grafts could revascularize 293 out of 310 vessels (94.5%) in the LAD approximately Diagonal region, 83 out of 113 (73.4%) in the distal RCA or Cx region. There were no cardiac events responsible for the arterial grafts in the follow up period. CONCLUSION In light of our experience, multiple revascularization with in situ arterial sequential grafts is feasible. Aggressive application of this technique provides patients requiring multiple coronary revascularization with favorable long-term results.