© 2000 American Heart Association, Inc.
Integrative Physiology |
Dephosphorylation and Intracellular Redistribution of Ventricular Connexin43 During Electrical Uncoupling Induced by Ischemia
From the Departments of Medicine, Surgery, Pediatrics and Pathology, and the Center for Cardiovascular Research, Washington University School of Medicine, St. Louis, Mo, and the Department of Physiology, University of Bern, Bern, Switzerland. Dr Beyer’s current address is Section of Pediatric Hematology/Oncology, University of Chicago Children’s Hospital, Chicago, Ill.
Correspondence to Jeffrey E. Saffitz, MD, PhD, Washington University School of Medicine, Department of Pathology, Box 8118, 660 S Euclid Ave, St. Louis, MO 63110. E-mail saffitz{at}pathology.wustl.edu
Abstract—Electrical uncoupling at gap junctions during acute myocardial ischemia contributes to conduction abnormalities and reentrant arrhythmias. Increased levels of intracellular Ca2+ and H+ and accumulation of amphipathic lipid metabolites during ischemia promote uncoupling, but other mechanisms may play a role. We tested the hypothesis that uncoupling induced by acute ischemia is associated with changes in phosphorylation of the major cardiac gap junction protein, connexin43 (Cx43). Adult rat hearts perfused on a Langendorff apparatus were subjected to ischemia or ischemia/reperfusion. Changes in coupling were monitored by measuring whole-tissue resistance. Changes in the amount and distribution of phosphorylated and nonphosphorylated isoforms of Cx43 were measured by immunoblotting and confocal immunofluorescence microscopy using isoform-specific antibodies. In control hearts, virtually all Cx43 identified immunohistochemically at apparent intercellular junctions was phosphorylated. During ischemia, however, Cx43 underwent progressive dephosphorylation with a time course similar to that of electrical uncoupling. The total amount of Cx43 did not change, but progressive reduction in total Cx43 immunofluorescent signal and concomitant accumulation of nonphosphorylated Cx43 signal occurred at sites of intercellular junctions. Functional recovery during reperfusion was associated with increased levels of phosphorylated Cx43. These observations suggest that uncoupling induced by ischemia is associated with dephosphorylation of Cx43, accumulation of nonphosphorylated Cx43 within gap junctions, and translocation of Cx43 from gap junctions into intracellular pools.
Key Words: connexin43 • gap junctions • ischemia • uncoupling • phosphorylation • arrhythmias
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