NaV1.5 | sodium voltage-gated channel alpha subunit 5
Family:
Sodium channels
Subgroups:
NaV1.1-1.9
Topology:
Alpha subunits consist of four homologous domains (I-IV) with six transmembrane alpha helices (S1–S6) and a pore-forming loop. One a subunit may associate with 1 or 2 b subunits to make up the channel.
NaV1.5: Background Information
NaV1.5 is found primarily in cardiac muscle, where it mediates the fast influx of Na+-ions (INa) across the cell membrane, resulting in the fast depolarization phase of the cardiac action potential. As such, it plays a major role in impulse propagation through the heart.. NaV1.5 is a TTX-resistent sodium channel.
Gene:
SCN5A
Human Protein:
UniProt Q14524
Tissue:
Cardiac myocytes, uninnervated skeletal muscle, central neurons, gastrointestinal smooth muscle cells and Interstitial cells of Cajal
Function/ Application:
Myocardial conduction, generation of action potentials and cell excitability
Pathology:
Romano-Ward, Brugada syndrome, Jervell, Lange-Nielsen, Long QT syndrome (LQT3), pain, cancer, gastrointestinal: Irritable bowel syndrome
Accessory subunits:
b1, b2, b3, b4
Interaction:
β1, β2, β3, β4 subunit, syntrophin, NEDD4, NEDD4L, WWP2, calmodulin
Modulator:
Aconitine, veratridine, α-scorpion toxin, ATX-II, saxitoxin, tetrodotoxin, lidocaine
Assays:
Patch Clamp: whole cell, room temperature, State- and use-dependence
Particularities:
NaV channel analysis requires GigaOhm seals and a stable and low access resistance
Recommended Reviews:
International Union of Pharmacology. XLVII. Nomenclature and Structure-Function Relationships of Voltage-Gated Sodium Channels. Pharmacol Rev 57: 397–409, Catterall, et al. 2005
Data and Applications
NaV1.5 - Raw current traces and Current-Voltage Relationship
Port-a-Patch mini data and applications:
Cells were kindly provided by Charles River.
The image shows currents from an example cell and the corresponding current-voltage plot for NaV1.5 expressed in stably-transfected CHO-K1 cells.
NaV1.5 - Tetracaine concentration response curve
Port-a-Patch mini data and applications:
Cells were kindly provided by Charles River.
The image shows the timecourse of the experiment (top) where peak amplitude is plotted against time and example traces in control conditions and different concentrations of tetracaine are also shown in the insert. The concentration reponse curve for tetracaine is shown on the bottom for an average of 6 cells. The curve is fit with a Hill equation revealing an IC50 = 25.5 ± 5.7 µM (n = 6).
NaV-Late - Recorded from Cardiosight-S hiPSC-CMs
Port-a-Patch mini data and applications:
Cells were kindly provided by Nexel.
NaV-Late was induced by application of ATX II in Cardiosight-S® cells on the Port-a-Patch mini. Example traces are shown on the top with the response to the voltage ramp (CiPA protocol) shown in the inset. The timecourse of the experiment is shown on the bottom, with block by ranolazine.
NaV1.5 - Late Current Analysis using the CiPA Protocol
SyncroPatch 384/768 PE (a predecessor model of the SyncroPatch 384/768i) data and applications:
Cells were kindly provided by Charles River.
Screenshots of the PatchControl 384 software showing NaV1.5 current traces in response to the CiPA voltage step protocol, measured on the SyncroPatch 384PE using whole cell patch clamp methodology and single-hole chips. The NaV1.5 late current was activated by the application of 60 nM ATX-II. The IC50 value of Ranolazine of the late Sodium current current was determined as 40.4 µM.
Cardiac Ion Channels - Pharmacology of Sotalol
CardioExcyte 96 and SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) data and applications:
Cells were kindly provided by Charles River and Cellular Dynamics.
The image on the left hand side displays the results of the blocking effect of Sotalol on hERG. The result is in good agreement with manual patch clamp data (Crumb et al., 2016). The compound induced arrhythmia when iPSC-CM were exposed to a minimum concentration of 10 µM. Arrhytmic events were both detected in field potential recordings as well as in the impedance based contractility measurements.
Cardiac Ion Channels - Pharmacology of Vandetanib
CardioExcyte 96 and 
Patchliner data and applications:
Cells were kindly provided by Charles River and Cellular Dynamics.
The image on the left hand side displays the results of the blocking effect of Vandetanib on hERG, NaV1.5, CaV1.2 and KV4.3. The compound induced arrhythmia when iPSC-CM were exposed to a minimum concentration of 1 µM. Arrhytmic events were both detected in field potential recordings as well as in the impedance based contractility.
NaV1.5 - Analyzing iPSC-derived Cardiomyocytes
SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly supplied by Cellular Dynamics.
Inhibition of NaV1.5 currents in stem cell-derived cardiomyocytes (iCells) by lidocaine. Lidocaine concentrations: 6 µM, 62 µM and 620 µM. The obatined IC50 -value was 14 µM.
NaV1.5 - Blind Study using an Inactivation Protocol
SyncroPatch data (a predecessor model of SyncroPatch 384PE) and applications:
22 selected compounds were tested in a blind study on the SyncroPatch 96 using HEK 293 cells expressing hNav1.5 ion channels. The IC50-values were compared to manual patch clamp measurements, performed at the customer site.
NaV1.5 - Screening Online Analysis
SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Millipore.
The image shows the state dependent block at -130 mV (light blue) and -90 mV (dark blue) by the addition of increasing compound concentrations to the different recording wells. The large window shows the effect of increasing concentrations of Lidocaine (red box) on the hNav1.5 currents.
NaV1.5 - Lidocaine Dose Response
SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Cytomyx Millipore
Timecourse of the NaV1.5 peak currents in response to exposure to different lidocaine concentrations (0 μM, 1 μM, 10 μM, 100 μM, 0 μM). Time points at which the external solution was exchanged is marked by the red lines.
NaV1.5 - Inactivation Protocol
SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Cytomyx Millipore.
Shown are raw current responses of HEK293 cells expressing hNaV1.5 to a double (inactivation) pulse protocol.
NaV1.5 - Screening Mode
SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Millipore.
Compound effect on hNav1.5 was investigated. A two-pulse protocol (-90 mV / -130 mV) was used to establish if the different compounds blocked the Nav1.5 currents in a state-dependent manner.
The image shows raw data traces. Success rate for the experiment was 71% (>1 GOhm seal resistance), indicated by the green color.
The corresponding online analysis of this experiment is shown in the data set NaV1.5 - Screening Online Analysis.
NaV1.5 - Stable Access Resistance
Patchliner data and applications:
Cells were kindly provided by Millipore.
The I/V-characteristics of NaV1.5 currents (HEK293) are shown together with the repeated dose dependent block by TTX (lower panel). Five concentrations of TTX (0.3, 1, 3, 10, 30 μM) were applied, followed by washout with antagonist-free buffer and re-application of the same TTX concentrations.
NaV1.5 - Accurate Voltage Control
Port-a-Patch data and applications:
Cells stably transfected with human SCN5A were kindly provided by Millipore.
To perform recordings of fast events, such as the activation and inactivation of Na currents, it is essential to have accurate voltage control. The image shows currents and I/V characteristics of NaV1.5 expressed in HEK293 cells.
NaV1.5 - Current Voltage Relationship
SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) data and applications:
Cells were kindly provided by Millipore.
Borosilicate glass chips are used as the patch clamp substrate, ensuring excellent voltage clamp of the cell membrane and high quality seals. Voltage gated channels such as hNaV1.5 (HEK293) have been used to validate the system. This data example shows the I/V characteristics and the corresponding raw current traces of a single cell from a recording on the SyncroPatch 96.
NaV1.5 - Lidocaine Block
Patchliner data and applications:
Cells were kindly provided by Cytomyx/Millipore.
Full dose response curves at different holding potentials were recorded for each cell (hNav1.5 in HEK293). Currents were elicited by a 10 ms voltage step to 0 mV. Plotted are average peak currents as a function of holding potential and lidocaine concentration.
Application Notes
Cardiac Ion Channels - "High Throughput Screening of Cardiac Ion Channels"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) Patchliner CardioExcyte 96 application note 
(2.3 MB)
Cardiac Ion Channels - "Simultaneous Assessment of CiPA Stipulated Ion Channels on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) application note (1.3 MB)
Cells were kindly provided by Charles River.
Cardiomyocytes - "Combining automated patch clamp, impedance and EFP of hiPSC-CMs"
CardioExcyte 96 SyncroPatch 3984PE (a predecessor model of SyncroPatch 384i) Patchliner Application Note
Cells kindly provided by Takara-Clonetech.
Cardiomyocytes - "Impedance and EFP recordings of Pluricyte Cardiomyocytes on the CardioExcyte 96"
CardioExcyte 96 Application Note (1.3 MB)
Cells were kindly provided by Ncardia.
NaV1.5 - "Characterization of CreaCell's hNaV1.5 (A-0822) on Nanion's Patchliner"
Patchliner application note: (0.9 MB)
Cells were kindly provided by CreaCell.
NaV1.5 - "High Throughput Pharmacology of NaV1.5 Channels on Nanion's SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) application note (1.6 MB)
Cells were kindly provided by Millipore.
NaV1.5 - "Pharmacology of hNaV1.5 recorded on Nanion's Patchliner"
Patchliner application note: (0.3 MB)
Cells were kindly provided by Millipore.
NaV1.5-Late - "INa-Late recorded from CHO cells and hiPSC-CMs on Nanion´s Patchliner"
Patchliner application note: (0.5 MB)
Cells were kindly provided by Charles River.
iCell® Cardiomyocytes2 were kindly provided by Fujifilm Cellular Dynamics International.
Webinars
Publications
2019 - High-throughput reclassification of SCN5A variants
SyncroPatch 384PE (a predecessor model of the SyncroPatch 384i) Pre-publication in bioRxiv Biology (2019)
Authors:
Glazer A.M., Wada Y., Li B., Muhammad A., Kalash O.R., O’Neill M.J., Shields T., Hall L., Short L., Blair M.A., Kroncke B.M., Capra J.A., Roden D.M
2019 - Electrophysiological evaluation of pentamidine and 17-AAG in human stem cell-derived cardiomyocytes for safety assessment
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) article in European Journal of Pharmacology (2019)
Authors:
Asahi Y., Nomura F., Abe Y., Doi M., Sakakura T., Takasuna K., Yasuda K.
2018 - The opioid oxycodone use‐dependently inhibits the cardiac sodium channel Nav1.5
CardioExcyte 96 publication in British Journal of Pharmacology (2018)
Authors:
Meents J.E., Juhasz K., Stölzle-Feix S., Peuckmann-Post V., Rolke R. Lampert A.
2018 - Multifocal atrial and ventricular premature contractions with an increased risk of dilated cardiomyopathy caused by a Nav1.5 gain-of-function mutation (G213D)
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) -related publication in International Journal of Cardiology (2018)
Authors:
Calloe K., Broendberg A.K., Christensen A.H., Pedersen L.N., Olesen M.S., de los Angeles Tejada M., Friis S., Thomsen M.B., Bundgaard H., Jensen H.K.
2017 - L-Type Calcium Channel Inhibition Contributes to the Proarrhythmic Effects of Aconitine in Human Cardiomyocytes
Patchliner publication in PLoS ONE (2017)
Authors:
Wu J., Wang X., Chung Y.Y. Koh C.H. Liu Z., Guo H., Yuan Q., Wang C., Su S., Wei H.
2017 - High-throughput electrophysiological assays for voltage gated ion channels using SyncroPatch 768PE
SyncroPatch 768PE (a predecessor model of SyncroPatch 384/768i) publication in PLoS One (2017)
Authors:
Li T, Lu G, Chiang E.Y., Chernov-Rogan T., Grogan J.L., Chen J.
2017 - Correlation between human ether-a-go-go related gene channel inhibition and action potential prolongation
Patchliner publication in British Journal of Pharmacology (2017)
Authors:
Saxena P., Hortigon‐Vinagre M.P., Beyl S.,Baburin I., Andranovits S., Iqbal S.M., Costa A., IJzerman A.P., Kügler P., Timin E., Smith G.L., Hering S.
2017 - A mutant of the Buthus martensii Karsch antitumor-analgesic peptide exhibits reduced inhibition to hNav1.4 and hNav1.5 channels while retaining analgesic activity
Port-a-Patch publication in Journal of Biological Chemistry (2017)
Authors:
Xu Y., Meng X., Hou X., Sun J., Kong X., Sun Y., Liu Z., Ma Y., Niu Y., Song Y., Cui Y., Zhao M., Zhang J.
2016 - Use-dependent Block of Human Cardiac Sodium Channels by GS967
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in Molecular Pharmacology (2016)
Authors:
Potet F., Vanoye C.G., George Jr. A.L.
2016 - Automated Patch Clamp Meets High-Throughput Screening: 384 Cells Recorded in Parallel on a Planar Patch Clamp Module
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in Journal of Lab Automation (2016)
Authors:
Obergrussberger A., Brüggemann A., Goetze T.A., Rapedius M., Haarmann C., Rinke I., Becker N., Oka T., Ohtsuki A., Stengel T., Vogel M., Steindl J., Mueller M., Stiehler J., George M., Fertig N.
2016 - Automated Electrophysiological and Pharmacological Evaluation of Human Pluripotent Stem Cell-Derived Cardiomyocytes
Patchliner publication in Stem Cells and Development (2016)
Authors:
Rajamohan D., Kalra S., Hoang M.D., George V., Staniforth A., Russell H., Yang X., Denning C.
2015 - The Functional Property Changes of Muscular Nav1.4 and Cardiac Nav1.5 Induced by Scorpion Toxin BmK AGP-SYPU1 Mutants Y42F and Y5F
Port-a-Patch publication in Biochemistry (2015)
Authors:
Meng x., Xu Y., Zhao M., Wang F., Ma Y., Jin Y., Liu Y., Song Y., Zhang J.
2015 - Scalable Electrophysiological Investigation of iPS Cell-Derived Cardiomyocytes Obtained by a Lentiviral Purification Strategy
Patchliner publication in Journal of Clinical Medicine (2015)
Authors:
Friedrichs S., Malan D., Voss Y., Sasse P.
2015 - Novel screening techniques for ion channel targeting drugs
Patchliner, SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) and CardioExcyte 96 publication in Channels (2015)
Authors:
Obergrussberger A., Stölzle-Feix S., Becker N., Brüggemann A., Fertig N., Möller C.
2014 - New strategies in ion channel screening for drug discovery: are there ways to improve its productivity?
SyncroPatch 384PE (a predecessor model of SyncroPatch 384i) publication in Journal of Laboratory Automation (2014)
Authors:
Farre C., Fertig N.
2012 - Isolation, characterization and total regioselective synthesis of the novel μO-conotoxin MfVIA from Conus magnificus that targets voltage-gated sodium channels
Port-a-Patch publication in Biochemical Pharmacology (2012)
Authors:
Vetter I., Dekan Z., Knapp O., Adams D.J., Alewood P.F., Lewis R.J.
2011 - State-of-the-art automated patch clamp devices: heat activation, action potentials, and high throughput in ion channel screening
Port-a-Patch, Patchliner and SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) publication in Frontiers in Pharmacology (2011)
Authors:
Stoelzle S., Obergrussberger A., Brüggemann A., Haarmann C., George M., Kettenhofen R., Fertig N.
2010 - Propranolol blocks cardiac and neuronal voltage-gated sodium channels
Patchliner publication in Frontiers in Pharmacology (2010)
Authors:
Wang D.W., Mistry A.M., Kahlig K.M., Kearney J.A, Xiang J., George Jr. A.L.
2009 - Port-a-Patch and Patchliner: High fidelity electrophysiology for secondary screening and safety pharmacology
Port-a-Patch and Patchliner publication in Combinatorial Chemistry & High Throughput Screening (2009)
Authors:
Farre C., Haythornthwaite A., Haarmann C., Stoelzle S., Kreir M., George M., Brüggemann A., Fertig N.
2007 - Automated ion channel screening: patch clamping made easy
Port-a-Patch and Patchliner publication in Expert Opinion Therapeutic Targets (2007)
Authors:
Farre C., Stoelzle S., Haarman C., George M., Brueggemann A., Fertig N.
2006 - Microchip technology for automated and parallel patch clamp recording
Port-a-Patch and Patchliner publication in Small Journal (2006)
Authors:
Brüggemann A., Stoelzle S., George M., Behrends J.C., Fertig N.