© 1997 American Heart Association, Inc.
Articles |
Antisense Oligodeoxynucleotides Directed Against Kv1.5 mRNA Specifically Inhibit Ultrarapid Delayed Rectifier K+ Current in Cultured Adult Human Atrial Myocytes
From the Department of Medicine and Research Center (J.F., G.-R.L., Z.W., S.N.), Montreal (Canada) Heart Institute; the Department of Medicine (G.-R.L., Z.W., S.N.), University of Montreal (Canada); the Department of Pharmacology and Therapeutics, McGill University (S.N.), Montreal, Quebec, Canada; and the Rammelkamp Center for Research (B.W.), MetroHealth Campus, Case Western Reserve University, Cleveland, Ohio.
Correspondence to Stanley Nattel, MD, Montreal Heart Institute, 5000 Belanger St East, Montreal, Quebec H1T 1C8, Canada.
Abstract Several cloned K+ channel subunits are
candidates to underlie macroscopic currents in the human heart, but
direct evidence bearing on their role is lacking. The Kv1.5
K+ channel subunit has been suggested to play a potential
role in human cardiac ultrarapid delayed rectifier (IKur)
and transient outward (Ito) currents. To evaluate the role
of proteins encoded by the Kv1.5 gene, we incubated cultured human
atrial myocytes for 48 hours in medium containing antisense
phosphorothioate oligodeoxynucleotides directed against octodecameric
segments of the Kv1.5 mRNA coding sequence, the same concentration of
homologous oligodeoxynucleotides with four mismatch mutations, or
vehicle (control group). Cells exposed to antisense showed a highly
significant (
50%) reduction in IKur, whether measured
by step current at the end of a 400-millisecond depolarizing pulse,
tail current at -20 mV, or current sensitive to a concentration of
4-aminopyridine (50 µmol/L) that is highly selective for
IKur, compared with control cells or cells exposed to
mismatch oligodeoxynucleo-tides. In contrast, Ito was
not different among the three experimental groups. When cultured human
ventricular myocytes were exposed to Kv1.5 antisense
oligodeoxynucleotides with the same controls, no changes occurred in
either Ito or the sustained current at the end of a
depolarizing pulse. We conclude that Kv1.5 channel subunits are
essential to the expression of IKur and do not play a role
in Ito in cultured human atrial myocytes. These studies
provide the first direct evidence with an antisense approach for the
equivalence between a macroscopic cardiac K+ current and a
cloned K+ channel subunit and offer insights into the
molecular electrophysiology of the human heart.
Key Words: K+ channel • antiarrhythmic drug • heart electrophysiology • cardiac action potential • molecular genetics
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B. London, W. Guo, X.-h. Pan, J. S. Lee, V. Shusterman, C. J. Rocco, D. A. Logothetis, J. M. Nerbonne, and J. A. Hill Targeted Replacement of Kv1.5 in the Mouse Leads to Loss of the 4-Aminopyridine-Sensitive Component of IK,slow and Resistance to Drug-Induced QT Prolongation Circ. Res., May 11, 2001; 88(9): 940 - 946. [Abstract] [Full Text] [PDF]
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V. Trepanier-Boulay, C. St-Michel, A. Tremblay, and C. Fiset Gender-Based Differences in Cardiac Repolarization in Mouse Ventricle Circ. Res., August 31, 2001; 89(5): 437 - 444. [Abstract] [Full Text] [PDF]
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