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 - Late Current Analysis using the CiPA Protocol

CiPA PE NaV1 5 Pharmacology late currenticon sp96   SyncroPatch 384/768 PE 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

CiPA PE CE Pharmacology SotalolIcon CE    CardioExcyte 96 and   icon sp96   SyncroPatch 384PE 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

CiPA PE CE Pharmacology VandetanibIcon CE    CardioExcyte 96 and   icon pl   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

0NavLidocaineScreenicon sp96   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

NaInactivation

icon sp96   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 NaOA small

icon sp96   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

NaLidocaine

icon sp96   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

NaInactivation

icon sp96   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 NaTrace small

icon sp96   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

p35 2 VoltContricon pl   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

p14 4 Nav

icon pap   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

NaStrom IV

icon sp96   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

application hnav15 3 small

icon pl   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"

icon sp96   SyncroPatch 384PE   icon pl   Patchliner   Icon CE   CardioExcyte 96 application note   logo pdf   (2.3 MB)

Cardiac Ion Channels - "Simultaneous Assessment of CiPA Stipulated Ion Channels on the SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE application note   logo pdf   (1.3 MB)
Cells were kindly provided by Charles River.

Cardiomyocytes - "Combining automated patch clamp, impedance and EFP of hiPSC-CMs"

Icon CE   CardioExcyte 96   icon sp96   SyncroPatch 3984PE   icon pl   Patchliner Application Note 
Cells kindly provided by Takara-Clonetech.

Cardiomyocytes - "Impedance and EFP recordings of Pluricyte Cardiomyocytes on the CardioExcyte 96"

Icon CE   CardioExcyte 96 Application Note   logo pdf   (1.3 MB)
Cells were kindly provided by Ncardia.

NaV1.5 - "Characterization of CreaCell's hNaV1.5 (A-0822) on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.9 MB)
Cells were kindly provided by CreaCell.

NaV1.5 - "High Throughput Pharmacology of NaV1.5 Channels on Nanion's SyncroPatch 384PE"

icon sp96   SyncroPatch 384PE application note   logo pdf   (1.9 MB)
Cells were kindly provided by Millipore.

NaV1.5 - "Pharmacology of hNaV1.5 recorded on Nanion's Patchliner"

icon pl   Patchliner application note:   logo pdf   (0.3 MB)
Cells were kindly provided by Millipore.

Webinars

03.11.2016 | External Webinar: Accelerating Ion Channel Characterization and New Drug Candidate Identification

icon sp96   SyncroPatch 384PE

This webinar will show high-throughput functional annotation of human ion channel variants associated with excitation disorders will be described along with use of the Syncropatch 384PE to measure subtype selective activation of KV7 potassium channels as well as inhibition of voltage gated sodium channels like NaV1.7, NaV1.1, and NaV1.5.
Organisation: Icagen Inc.

Publications

2018 - The opioid oxycodone use‐dependently inhibits the cardiac sodium channel Nav1.5

Icon CE   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)

icon sp96   SyncroPatch 384PE-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

icon pl  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

icon sp96  SyncroPatch 768PE 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

icon pl   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

icon pap   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

icon sp96  SyncroPatch 384PE 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

icon sp96  SyncroPatch 384PE 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

icon pl  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

icon pap  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

icon pl  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

icon pl  Patchliner,   icon sp96   SyncroPatch 384PE and   Icon CE   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?

icon sp96  SyncroPatch 384PE 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

icon pap   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

icon pap   Port-a-Patch,   icon pl  Patchliner and   icon sp96   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

icon pl  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

icon pap  Port-a-Patch and   icon pl   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

icon pap  Port-a-Patch and   icon pl   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

icon pap  Port-a-Patch and   icon pl   Patchliner publication in Small Journal (2006)

Authors: 
Brüggemann A., Stoelzle S., George M., Behrends J.C., Fertig N.

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