© 1999 American Heart Association, Inc.
Integrative Physiology |
Attenuation of the Slow Component of Delayed Rectification, Action Potential Prolongation, and Triggered Activity in Mice Expressing a Dominant-Negative Kv2 
Subunit
From the Department of Molecular Biology and Pharmacology, Washington University Medical School, St. Louis, Mo.
Correspondence to Jeanne M. Nerbonne, Department of Molecular Biology and Pharmacology, Washington University Medical School, 660 S Euclid Ave, St. Louis, MO 63110. E-mail jnerbonn{at}pharmdec.wustl.edu
Abstract—An in vivo experimental
strategy, involving cardiac-specific expression of a mutant Kv 2.1
subunit that functions as a dominant negative, was exploited in studies
focused on exploring the role of members of the Kv2 subfamily of
pore-forming (
) subunits in the generation of functional
voltage-gated K+ channels in the mammalian heart. A mutant
Kv2.1 subunit (Kv2.1N216) was designed to produce a truncated
protein containing the intracellular N terminus, the S1
membrane–spanning domain, and a portion of the S1/S2 loop. The
truncated Kv2.1N216 was epitope tagged at the C terminus with the
8–amino acid FLAG peptide to generate Kv2.1N216FLAG. No ionic currents
are detected on expression of Kv2.1N216FLAG in HEK-293 cells, although
coexpression of this construct with wild-type Kv2.1 markedly reduced
the amplitudes of Kv2.1-induced currents. Using the -myosin heavy
chain promoter to direct cardiac specific expression of the transgene,
2 lines of Kv2.1N216FLAG-expressing transgenic mice were generated.
Electrophysiological recordings from
ventricular myocytes isolated from these animals revealed
that IK, slow is selectively reduced. The
attenuation of IK, slow is accompanied by
marked action potential prolongation, and, occasionally, spontaneous
triggered activity (apparently induced by early afterdepolarizations)
is observed. The time constant of inactivation of
IK, slow in Kv2.1N216FLAG-expressing cells
(mean±SEM=830±103 ms; n=17) is accelerated compared with the time
constant of IK, slow inactivation
(mean±SEM=1147±57 ms; n=25) in nontransgenic cells. In addition,
unlike IK, slow in wild-type cells, the
component of IK, slow remaining in the
Kv2.1N216FLAG-expressing cells is insensitive to 25 mmol/L
tetraethylammonium. Taken together, these
observations suggest that there are 2 distinct components of
IK, slow in mouse ventricular
myocytes and that Kv2 subunits underlie the more slowly
inactivating, tetraethylammonium-sensitive
component of IK, slow. In vivo telemetric
recordings also reveal marked QT prolongation,
consistent with a defect in ventricular
repolarization, in Kv2.1N216FLAG-expressing transgenic mice.
Key Words: transgenic mice • ventricle • action potential • triggered activity • arrhythmia
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