<h3>Introduction</h3> Brugada syndrome is associated with ventricular tachycardia originating in the right ventricle (RV); this has been attributed to either depolarisation abnormalities or increased repolarisation heterogeneities. We have used a heterozygotic <i>Scn5a+/−</i> murine model to investigate the underlying mechanisms for the predisposition of the RV to arrhythmias. <h3>Methods and Results</h3> Na<sub>v</sub>1.5 mRNA and protein expression were lower in <i>Scn5a+/−</i> than wild-type (WT) hearts, with a further reduction in the RV compared to left ventricle (LV) (Abstract E figure 1A,B, n=4, significant differences: * = WT vs <i>Scn5a+/−</i>; # = LV vs RV). There were higher expression levels of K<sub>v</sub>4.2, K<sub>v</sub>4.3 and KChIP<sub>2</sub> in RV than LV in both groups. Action potential (AP) upstroke velocity was decreased in <i>Scn5a+/−</i> (RV: 59.43±2.70 V/s to 30.26±4.03 V/s, p<0.0001, n=20), and furthermore was smaller in RV than LV. AP durations were smallest in the RV of <i>Scn5a+/−</i> myocytes. RV transient outward current density (<i>I<sub>to</sub></i>) was greater than LV in both WT and <i>Scn5a+/−</i> (Abstract E figure 1C, n=17), with similar voltage dependence of activation. Time constants of inactivation were larger in RV than LV, and voltage dependence of inactivation was shifted to more negative values in RV compared to LV, but to more positive values in <i>Scn5a+/−</i> compared to WT. Maximum Na<sup>+</sup> current density (<i>I<sub>Na</sub></i>) was decreased in <i>Scn5a+/−</i>, with a further reduction in the RV (Abstract E figure 2A–C, n=17). Voltage dependence of activation was unchanged, but inactivation was shifted to more negative values in <i>Scn5a+/−</i>. Maximum persistent Na<sup>+</sup> current density (<i>I<sub>pNa</sub></i>) was decreased in a similar pattern to <i>I<sub>Na</sub></i> (RV: −0.30±0.03 pA/pF, n=15 vs −0.17±0.02 pA/pF, n=22, p=0.0009). <h3>Conclusion</h3> Our findings show preferential upregulation of the single Scn5a gene in the LV of the <i>Scn5a+/−</i> mice compared to RV. The reduced expression of Na<sup>+</sup> channels in RV leads to smaller <i>I<sub>Na</sub></i>, resulting in slowed conduction, and smaller <i>I<sub>pNa</sub></i>, which in combination with increased <i>I<sub>to</sub></i>, results in shorter AP durations and greater heterogeneity of repolarisation, thus suggesting arrhythmogenesis may be initiated by both abnormal depolarisation and repolarisation in the RV of <i>Scn5a+/−</i> hearts. Insight into the molecular mechanisms of arrhythmias could prove crucial in planning possible new pharmacological therapies for a disease where the mainstay of treatment is cardioverter-defibrillator implantation.