AUTHOR=Rougier Jean-Sébastien , Essers Maria C. , Gillet Ludovic , Guichard Sabrina , Sonntag Stephan , Shmerling Doron , Abriel Hugues 

TITLE=A Distinct Pool of Nav1.5 Channels at the Lateral Membrane of Murine Ventricular Cardiomyocytes

JOURNAL=Frontiers in Physiology

VOLUME=10

YEAR=2019

URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2019.00834

DOI=10.3389/fphys.2019.00834

ISSN=1664-042X

ABSTRACT=<p><bold>Background:</bold> In cardiac ventricular muscle cells, the presence of voltage-gated sodium channels Na<sub>v</sub>1.5 at the lateral membrane depends in part on the interaction between the dystrophin–syntrophin complex and the Na<sub>v</sub>1.5 C-terminal PDZ-domain-binding sequence Ser-Ile-Val (SIV motif). α1-Syntrophin, a PDZ-domain adaptor protein, mediates the interaction between Na<sub>v</sub>1.5 and dystrophin at the lateral membrane of cardiac cells. Using the cell-attached patch-clamp approach on cardiomyocytes expressing Na<sub>v</sub>1.5 in which the SIV motif is deleted (ΔSIV), sodium current (I<sub>Na</sub>) recordings from the lateral membrane revealed a SIV-motif-independent I<sub>Na</sub>. Since immunostaining has suggested that Na<sub>v</sub>1.5 is expressed in transverse (T-) tubules, this remaining I<sub>Na</sub> might be carried by channels in the T-tubules. Of note, a recent study using heterologous expression systems showed that α1-syntrophin also interacts with the Na<sub>v</sub>1.5 N-terminus, which may explain the SIV-motif independent I<sub>Na</sub> at the lateral membrane of cardiomyocytes.</p><p><bold>Aim:</bold> To address the role of α1-syntrophin in regulating the I<sub>Na</sub> at the lateral membrane of cardiac cells.</p><p><bold>Methods and Results:</bold> Patch-clamp experiments in cell-attached configuration were performed on the lateral membranes of wild-type, α1-syntrophin knockdown, and ΔSIV ventricular mouse cardiomyocytes. Compared to wild-type, a reduction of the lateral I<sub>Na</sub> was observed in myocytes from α1-syntrophin knockdown hearts. Similar to ΔSIV myocytes, a remaining I<sub>Na</sub> was still recorded. In addition, cell-attached I<sub>Na</sub> recordings from lateral membrane did not differ significantly between non-detubulated and detubulated ΔSIV cardiomyocytes. Lastly, we obtained evidence suggesting that cell-attached patch-clamp experiments on the lateral membrane cannot record currents carried by channels in T-tubules such as calcium channels.</p><p><bold>Conclusion:</bold> Altogether, these results suggest the presence of a sub-pool of sodium channels at the lateral membrane of cardiomyocytes that is independent of α1-syntrophin and the PDZ-binding motif of Na<sub>v</sub>1.5, located in membrane domains outside of T-tubules. The question of a T-tubular pool of Na<sub>v</sub>1.5 channels, however, remains open.</p>