title: Ankyrin-G coordinates intercalated disc signaling platform to regulate cardiac excitability in vivo.
abstract: BACKGROUND
  Nav1.5 (SCN5A) is the primary cardiac voltage-gated Nav channel. Nav1.5 is critical for cardiac excitability and conduction, and human SCN5A mutations cause sinus node dysfunction, atrial fibrillation, conductional abnormalities, and ventricular arrhythmias. Further, defects in Nav1.5 regulation are linked with malignant arrhythmias associated with human heart failure. Consequently, therapies to target select Nav1.5 properties have remained at the forefront of cardiovascular medicine. However, despite years of investigation, the fundamental pathways governing Nav1.5 membrane targeting, assembly, and regulation are still largely undefined.
  
  
  OBJECTIVE
  Define the in vivo mechanisms underlying Nav1.5 membrane regulation.
  
  
  RESULTS
  Here, we define the molecular basis of an Nav channel regulatory platform in heart. Using new cardiac-selective ankyrin-G(-/-) mice (conditional knock-out mouse), we report that ankyrin-G targets Nav1.5 and its regulatory protein calcium/calmodulin-dependent kinase II to the intercalated disc. Mechanistically, βIV-spectrin is requisite for ankyrin-dependent targeting of calcium/calmodulin-dependent kinase II-δ; however, βIV-spectrin is not essential for ankyrin-G expression. Ankyrin-G conditional knock-out mouse myocytes display decreased Nav1.5 expression/membrane localization and reduced INa associated with pronounced bradycardia, conduction abnormalities, and ventricular arrhythmia in response to Nav channel antagonists. Moreover, we report that ankyrin-G links Nav channels with broader intercalated disc signaling/structural nodes, as ankyrin-G loss results in reorganization of plakophilin-2 and lethal arrhythmias in response to β-adrenergic stimulation.
  
  
  CONCLUSIONS
  Our findings provide the first in vivo data for the molecular pathway required for intercalated disc Nav1.5 targeting/regulation in heart. Further, these new data identify the basis of an in vivo cellular platform critical for membrane recruitment and regulation of Nav1.5.
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Michael A
authors.initials: MA
authors.lastname: Makara
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Jerry
authors.initials: J
authors.lastname: Curran
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Sean C
authors.initials: SC
authors.lastname: Little
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Hassan
authors.initials: H
authors.lastname: Musa
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Iuliia
authors.initials: I
authors.lastname: Polina
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Sakima A
authors.initials: SA
authors.lastname: Smith
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Wright
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Sathya D
authors.initials: SD
authors.lastname: Unudurthi
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Jed
authors.initials: J
authors.lastname: Snyder
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Vann
authors.initials: V
authors.lastname: Bennett
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.).
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Hund
authors.affiliation: From The Dorothy M. Davis Heart & Lung Research Institute (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., S.D.U., J.S., T.J.H., P.J.M. ), Departments of Internal Medicine (S.A.S., P.J.M.), and Physiology and Cell Biology (M.A.M., J.C., S.C.L., H.M., I.P., S.A.S., P.J.W., P.J.M.), The Ohio State University Wexner Medical Center, Columbus; Department of Biomedical Engineering, The Ohio State University College of Engineering, Columbus (S.D.U., J.S., T.J.H.); and Howard Hughes Medical Institute, Department of Biochemistry, Duke University Medical Center, Durham, NC (V.B.). peter.mohler@osumc.edu.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Mohler
mesh_terms: D000200:Action Potentials; D000818:Animals; D017487:Ankyrins / Q000235:genetics / Q000378:metabolism*; D001145:Arrhythmias, Cardiac / Q000378:metabolism* / Q000503:physiopathology; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D062554:NAV1.5 Voltage-Gated Sodium Channel / Q000235:genetics / Q000378:metabolism*; D051191:Plakophilins / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D015398:Signal Transduction; D026941:Sodium Channel Blockers / Q000494:pharmacology; D013049:Spectrin / Q000378:metabolism
chemical_list: C094681:Ank3 protein, mouse; D017487:Ankyrins; D062554:NAV1.5 Voltage-Gated Sodium Channel; D051191:Plakophilins; C568321:Scn5a protein, mouse; D026941:Sodium Channel Blockers; D013049:Spectrin; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2
keywords: Nav1.5; ankyrin; arrhythmia (mechanisms); cell biology; mouse mutant; protein trafficking; sodium channels; targeting