Late Na currents affected by alpha subunit isoform and beta1 subunit co-expression in HEK293 cells.
Peak Na current underlies excitability and conduction in the heart, and late non-inactivating or slowly inactivating Na current plays a role in action potential duration. We hypothesized that different alpha subunit isoforms or beta1 subunit co-expression might affect late Na current. The human Na channel alpha subunits hNa(v)1.5 (hH1a) and hNa(v)1.4 (hSkM1) were transfected with and without the hNa(V)beta1 (beta1) subunit in HEK293 cells and studied by whole cell patch clamp. The inactivation relationship for hH1a was 28mV negative to that for hSkM1, and beta1 shifted the midpoint positively by 22mV for hH1a and 8mV for hSkM1. When pre-pulse duration was varied from 10ms to 10s, "steady-state" was approached more slowly for hH1a. beta1 caused hH1a but not hSkM1 to reach "steady-state" earlier. Both isoforms showed two recovery components but hH1a showed a "cardiac phenotype" with a smaller slow component that was unaffected by beta1. The amplitude of a late current (at 750ms) was significantly greater for hH1a than hSkM1, but beta1 decreased late current for hH1a and eliminated the difference. Under the study conditions the alpha subunit isoforms have distinct functional phenotypes and co-expression with beta1 tends to diminish these distinctions. These properties may provide mechanisms for regional and transmural distribution of late Na current and late Na current amplitudes during development and in disease states.
Valdivia
Carmen R
CR
Departments of Medicine and Physiology, University of Wisconsin, 600 Highland Avenue, Madison, WI 53792, USA.
Nagatomo
Toshihisa
T
Makielski
Jonathan C
JC
D006801:Humans; D007700:Kinetics; D062554:NAV1.5 Voltage-Gated Sodium Channel; D018408:Patch-Clamp Techniques; D020033:Protein Isoforms / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels / Q000378:metabolism*
D062554:NAV1.5 Voltage-Gated Sodium Channel; D020033:Protein Isoforms; C568320:SCN5A protein, human; D015222:Sodium Channels; D012964:Sodium