Direct cytosolic communication between cells is mediated by an aggregate of intercellular channels in the plasma membrane called the gap junction (GJ). Remodeling of connexin43 (Cx43) GJs has been linked to cardiac arrhythmias. Work accomplished during the previous period of this RO1 identified a biological function of interaction between Cx43 and the actin-binding protein Zonula Occludens (ZO)-1 in GJ remodeling. In recent work, we have determined that a peptide known to inhibit Cx43/ZO-1 interaction, 1CT1, reduces the frequency of inducible arrhythmias following injury to the left ventricle. Preliminary data from work performed with 1CT1 in vitro, indicates that the transition from free connexon channels in the membrane to paired connexons in intercellular channel aggregates underlies GJ remodeling. We hypothesize that ZO-1 regulates the rate at which free connexons in the membrane accrete to GJs-the "connexon switch".
The aims i n this renewal will test this hypothesis and its implications for GJ intercellular communication, membrane excitability, and a novel role for Cx43 in differential adhesion between myocytes and fibroblasts following myocardial infarction - all processes likely to impact susceptibility to re-entrant arrhythmia. The proposed experiments will quantitatively determine the role of Cx43/ZO-1 interaction in the "connexon switch" using methods including live cell imaging, fluorescent fusion proteins, loss-of-function mutants, whole cell patch clamp, single channel electrophysiology and biochemical assays. Implications of Cx43/ZO-1 interaction within the physiological framework of cardiac injury in vivo will be assessed by echocardiography, EKG telemetry, optical mapping of electrical activation and arrhythmia induction protocols. The data generated will broaden our understanding of fundamental mechanisms of Cx43 function and may translate to new therapies for the prevention and treatment of cardiac arrhythmia.

Public Health Relevance

Injury and scarring of the heart following myocardial infarction (heart attack) is one the most frequent causes of sudden death in the USA. This project will deliver new insights on the function of a protein essential to a stable heartbeat called connexin43. The project aims to provide novel approaches to the prevention and treatment of heart attacks.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
7R01HL056728-13
Application #
8438508
Study Section
Special Emphasis Panel (ZRG1-CVRS-E (02))
Program Officer
Krull, Holly
Project Start
2010-04-01
Project End
2014-05-31
Budget Start
2013-06-21
Budget End
2014-05-31
Support Year
13
Fiscal Year
2013
Total Cost
$341,413
Indirect Cost
$129,355
Name
Virginia Polytechnic Institute and State University
Department
None
Type
Organized Research Units
DUNS #
003137015
City
Blacksburg
State
VA
Country
United States
Zip Code
24061
Kohl, Peter; Gourdie, Robert G (2014) Fibroblast-myocyte electrotonic coupling: does it occur in native cardiac tissue? J Mol Cell Cardiol 70:37-46
Veeraraghavan, Rengasayee; Poelzing, Steven; Gourdie, Robert G (2014) Intercellular electrical communication in the heart: a new, active role for the intercalated disk. Cell Commun Adhes 21:161-7
Veeraraghavan, Rengasayee; Gourdie, Robert G; Poelzing, Steven (2014) Mechanisms of cardiac conduction: a history of revisions. Am J Physiol Heart Circ Physiol 306:H619-27
Rhett, J Matthew; Veeraraghavan, Rengasayee; Poelzing, Steven et al. (2013) The perinexus: sign-post on the path to a new model of cardiac conduction? Trends Cardiovasc Med 23:222-8
Rhett, Joshua Matthew; Gourdie, Robert G (2012) The perinexus: a new feature of Cx43 gap junction organization. Heart Rhythm 9:619-23
O'Quinn, Michael P; Palatinus, Joseph A; Harris, Brett S et al. (2011) A peptide mimetic of the connexin43 carboxyl terminus reduces gap junction remodeling and induced arrhythmia following ventricular injury. Circ Res 108:704-15
Palatinus, Joseph A; O'Quinn, Michael P; Barker, Ralph J et al. (2011) ZO-1 determines adherens and gap junction localization at intercalated disks. Am J Physiol Heart Circ Physiol 300:H583-94
Rhett, J Matthew; Jourdan, Jane; Gourdie, Robert G (2011) Connexin 43 connexon to gap junction transition is regulated by zonula occludens-1. Mol Biol Cell 22:1516-28
Palatinus, Joseph A; Rhett, Joshua M; Gourdie, Robert G (2011) Enhanced PKC? mediated phosphorylation of connexin43 at serine 368 by a carboxyl-terminal mimetic peptide is dependent on injury. Channels (Austin) 5:236-40
Palatinus, Joseph A; Rhett, J Matthew; Gourdie, Robert G (2010) Translational lessons from scarless healing of cutaneous wounds and regenerative repair of the myocardium. J Mol Cell Cardiol 48:550-7

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