title: Transport and regulation of the cardiac Na(+)-Ca2+ exchanger, NCX1. Comparison between Ca2+ and Ba2+.
abstract: Cardiac muscle fails to relax upon replacement of extracellular Ca2+ with Ba2+. Among the manifold consequences of this intervention, one major possibility is that Na(+)-Ba2+ exchange is inadequate to support normal relaxation. This could occur due to reduced transport rates of Na(+)-Ba2+ exchange and/or by failure of Ba2+ to activate the exchanger molecule at the high affinity regulatory Ca2+ binding site. In this study, we examined transport and regulatory properties for Na(+)-Ca2+ and Na(+)-Ba2+ exchange. Inward and outward Na(+)-Ca2+ or Na(+)-Ba2+ exchange currents were examined at 30 degrees C in giant membrane patches excised from Xenopus oocytes expressing the cloned cardiac Na(+)-Ca2+ exchanger, NCX1. When excised patches were exposed to either cytoplasmic Ca2+ or Ba2+, robust inward Na(+)-Ca2+ exchange currents were observed, whereas Na(+)-Ba2+ currents were absent or barely detectable. Similarly, outward currents were greatly reduced when pipette solutions contained Ba2+ rather than Ca2+. However, when solution temperature was elevated from 30 degrees C to 37 degrees C, a substantial increase in outward Na(+)-Ba2+ exchange currents was observed, but not so for inward currents. We also compared the relative abilities of Ca2+ and Ba2+ to activate outward Na(+)-Ca2+ exchange currents at the high affinity regulatory Ca2+ binding site. While Ba2+ was capable of activating the exchanger, it did so with a much lower affinity (KD approximately 10 microM) compared with Ca2+ (KD approximately 0.3 microM). Moreover, the efficiency of Ba2+ regulation of Na(+)-Ca2+ exchange is also diminished relative to Ca2+, supporting approximately 60% of maximal currents obtainable with Ca2+. Ba2+ is also much less effective at alleviating Na+i-induced inactivation of NCX1. These results indicate that the reduced ability of NCX1 to adequately exchange Na+ and Ba2+ contributes to failure of the relaxation process in the cardiac muscle.
authors.affiliation: Institute of Cardiovascular Sciences, University of Manitoba, St. Boniface General Hospital Research Centre, Winnipeg, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Trac
authors.forename: C
authors.initials: C
authors.lastname: Dyck
authors.forename: M
authors.initials: M
authors.lastname: Hnatowich
authors.forename: A
authors.initials: A
authors.lastname: Omelchenko
authors.forename: L V
authors.initials: LV
authors.lastname: Hryshko
mesh_terms: D000818:Animals; D001464:Barium / Q000378:metabolism / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism*; D004285:Dogs; D015536:Down-Regulation / Q000502:physiology; D004594:Electrophysiology; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000502:physiology; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011994:Recombinant Proteins / Q000378:metabolism; D019831:Sodium-Calcium Exchanger; D014982:Xenopus laevis
chemical_list: D002352:Carrier Proteins; D011994:Recombinant Proteins; D019831:Sodium-Calcium Exchanger; D001464:Barium; D002118:Calcium