title: A transgenic zebrafish model of a human cardiac sodium channel mutation exhibits bradycardia, conduction-system abnormalities and early death.
abstract: The recent exponential increase in human genetic studies due to the advances of next generation sequencing has generated unprecedented numbers of new gene variants. Determining which of these are causative of human disease is a major challenge. In-vitro studies and murine models have been used to study inherited cardiac arrhythmias but have several limitations. Zebrafish models provide an attractive alternative for modeling human heart disease due to similarities in cardiac electrophysiology and contraction, together with ease of genetic manipulation, external development and optical transparency. Although zebrafish cardiac mutants and morphants have been widely used to study loss and knockdown of zebrafish gene function, the phenotypic effects of human dominant-negative gene mutations expressed in transgenic zebrafish have not been evaluated. The aim of this study was to generate and characterize a transgenic zebrafish arrhythmia model harboring the pathogenic human cardiac sodium channel mutation SCN5A-D1275N, that has been robustly associated with a range of cardiac phenotypes, including conduction disease, sinus node dysfunction, atrial and ventricular arrhythmias, and dilated cardiomyopathy in humans and in mice. Stable transgenic fish with cardiac expression of human SCN5A were generated using Tol2-mediated transgenesis and cardiac phenotypes were analyzed using video microscopy and ECG. Here we show that transgenic zebrafish expressing the SCN5A-D1275N mutation, but not wild-type SCN5A, exhibit bradycardia, conduction-system abnormalities and premature death. We furthermore show that SCN5A-WT, and to a lesser degree SCN5A-D1275N, are able to compensate the loss of endogenous zebrafish cardiac sodium channels, indicating that the basic pathways, through which SCN5A acts, are conserved in teleosts. This proof-of-principle study suggests that zebrafish may be highly useful in vivo models to differentiate functional from benign human genetic variants in cardiac ion channel genes in a time- and cost-efficient manner. This article is part of a Special Issue entitled "Na(+) Regulation in Cardiac Myocytes".
authors.affiliation: Molecular Cardiology Division, Victor Chang Cardiac Research Institute, Darlinghurst, NSW 2010, Australia.
authors.forename: Inken G
authors.initials: IG
authors.lastname: Huttner
authors.forename: Gunjan
authors.initials: G
authors.lastname: Trivedi
authors.forename: Arie
authors.initials: A
authors.lastname: Jacoby
authors.forename: Stefan A
authors.initials: SA
authors.lastname: Mann
authors.forename: Jamie I
authors.initials: JI
authors.lastname: Vandenberg
authors.forename: Diane
authors.initials: D
authors.lastname: Fatkin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001919:Bradycardia / Q000235:genetics* / Q000503:physiopathology; D004195:Disease Models, Animal; D006329:Heart Conduction System / Q000002:abnormalities*; D006339:Heart Rate; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D062554:NAV1.5 Voltage-Gated Sodium Channel / Q000096:biosynthesis* / Q000235:genetics; D019683:Penetrance; D010641:Phenotype; D015027:Zebrafish / Q000235:genetics*
chemical_list: D062554:NAV1.5 Voltage-Gated Sodium Channel; C568320:SCN5A protein, human
keywords: AP; AV; CCD; Cardiac arrhythmia; Cardiac sodium channel; DCM; ECG; FS; Gene mutation; Na(+) channel; PCR; RT; Transgenesis; WT; Zebrafish; action potential; alpha-crystallin; atrio-ventricular; beats per minute; bpm; cardiac conduction disease; cardiac myosin light chain 2; cardiac sodium channel; cmlc2; days post fertilization; dilated cardiomyopathy; dpf; electrocardiogram; fractional shortening; polymerase chain reaction; reverse transcriptase; wild-type; α-cry