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Determination of the Relative Cell Surface and Total Expression of Recombinant Ion Channels Using Flow Cytometry
Published on: September 28, 2016
I(Ca(TTX)) channels are distinct from those generating the classical cardiac Na(+) current
Y Chen-Izu1, Q Sha, S R Shorofsky
1Department of Medicine, School of Medicine, University of Maryland, Baltimore, Maryland 21201, USA.
Calcium (Ca2+) is not required to demonstrate the tetrodotoxin-sensitive sodium (Na+) current component, I(Ca(TTX)). This study reveals two distinct sodium channel populations, not interconvertible, with differing calcium permeability.
Area of Science:
- Cardiovascular Physiology
- Ion Channel Biophysics
- Molecular Cardiology
Background:
- The tetrodotoxin-sensitive sodium current component, I(Ca(TTX)), is distinct from the main sodium current, I(Na).
- Previous hypotheses suggested I(Ca(TTX)) arises from I(Na) channels altered by specific ionic conditions (low Na+, high Ca2+).
Purpose of the Study:
- To investigate whether calcium (Ca2+) or sodium (Na+)-free conditions are necessary to observe I(Ca(TTX)).
- To determine if I(Na) channels can be converted into I(Ca(TTX)) channels.
Main Methods:
- Whole-cell patch-clamp electrophysiology on adult rat ventricular cells.
- Comparison of tetrodotoxin-blockable currents under varying ionic conditions (Cs+ vs. Ca2+).
- Heterologous expression of cardiac Na+ (H1) channels in HEK 293 cells.
Main Results:
- I(Ca(TTX)) can be demonstrated without Ca2+ or Na+-free conditions.
- Gating properties of Cs+ currents and I(Ca(TTX)) were similar after voltage correction, indicating non-interconvertibility.
- Heterologously expressed cardiac Na+ (H1) channels did not convert to I(Ca(TTX)) channels under proposed conversion conditions.
Conclusions:
- Two distinct, non-interconvertible Na+ channel populations exist in cardiac cells: one with low Ca2+ permeability and another with significant Ca2+ permeability.
- The proposed conversion mechanism of I(Na) to I(Ca(TTX)) channels is not supported by these findings.
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