title: Re-evaluation of the action potential upstroke velocity as a measure of the Na+ current in cardiac myocytes at physiological conditions.
abstract: BACKGROUND
  The SCN5A encoded sodium current (I(Na)) generates the action potential (AP) upstroke and is a major determinant of AP characteristics and AP propagation in cardiac myocytes. Unfortunately, in cardiac myocytes, investigation of kinetic properties of I(Na) with near-physiological ion concentrations and temperature is technically challenging due to the large amplitude and rapidly activating nature of I(Na), which may seriously hamper the quality of voltage control over the membrane. We hypothesized that the alternating voltage clamp-current clamp (VC/CC) technique might provide an alternative to traditional voltage clamp (VC) technique for the determination of I(Na) properties under physiological conditions.
  
  
  RESULTS
  We studied I(Na) under close-to-physiological conditions by VC technique in SCN5A cDNA-transfected HEK cells or by alternating VC/CC technique in both SCN5A cDNA-transfected HEK cells and rabbit left ventricular myocytes. In these experiments, peak I(Na) during a depolarizing VC step or maximal upstroke velocity, dV/dt(max), during VC/CC served as an indicator of available I(Na). In HEK cells, biophysical properties of I(Na), including current density, voltage dependent (in)activation, development of inactivation, and recovery from inactivation, were highly similar in VC and VC/CC experiments. As an application of the VC/CC technique we studied I(Na) in left ventricular myocytes isolated from control or failing rabbit hearts.
  
  
  CONCLUSIONS
  Our results demonstrate that the alternating VC/CC technique is a valuable experimental tool for I(Na) measurements under close-to-physiological conditions in cardiac myocytes.
authors.affiliation: Department of Anatomy, Embryology, and Physiology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Géza
authors.initials: G
authors.lastname: Berecki
authors.forename: Ronald
authors.initials: R
authors.lastname: Wilders
authors.forename: Berend
authors.initials: B
authors.lastname: de Jonge
authors.forename: Antoni C G
authors.initials: AC
authors.lastname: van Ginneken
authors.forename: Arie O
authors.initials: AO
authors.lastname: Verkerk
mesh_terms: D000200:Action Potentials; D000818:Animals; D001703:Biophysics / Q000379:methods; D002460:Cell Line; D018076:DNA, Complementary / Q000378:metabolism; D004594:Electrophysiology; D006321:Heart / Q000502:physiology; D006801:Humans; D007477:Ions; D008297:Male; D009206:Myocardium / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology*; D018408:Patch-Clamp Techniques; D011817:Rabbits; D012964:Sodium / Q000737:chemistry*; D015222:Sodium Channels / Q000737:chemistry
chemical_list: D018076:DNA, Complementary; D007477:Ions; D015222:Sodium Channels; D012964:Sodium