Inhibition of cardiac delayed rectifier K+ currents by an antisense oligodeoxynucleotide against IsK (minK) and over-expression of IsK mutant D77N in neonatal mouse hearts.
The IsK (minK or KCNE1) protein is known to co-assemble with the KvLQT1 (KCNQ1) protein to form a channel underlying the slowly activating delayed rectifier K+ current (IKs). Controversy remains as to whether the IsK protein assembles with ERG (the ether-a-go-go-related gene) products to form or modulate the channel-underlying the rapidly activating delayed rectifier K+ current (IKr). We investigated the effects of antisense oligodeoxynucleotides (AS-ODN) against IsK and its mutant D77N [which underlies a form of long QT syndrome (LQT5) in humans] on the delayed rectifier K+ current (IK) of neonatal mouse ventricular myocytes in primary culture. Patch-clamp experiments on these cells showed that IK consists of IKs and IKr. IK was not recorded from ventricular cells transfected with AS-ODN, while it was recorded from cells transfected with the corresponding sense oligodeoxynucleotides (S-ODN). IK was not recorded from cells transfected with the D77N mutant, and the action potential duration was much longer than in cells transfected with wild-type IsK. Furthermore, HERG could not induce currents in COS-1 cells co-expressed with the D77N mutant and HERG (the human form of ERG). These results indicate that the IsK protein associates with both KvLQT1 and ERG products to modulate IKr and IKs in cardiac myocytes.
Ohyama
H
H
Department of Physiology, Kansai Medical University, Moriguchi, Osaka, Japan.
Kajita
H
H
Omori
K
K
Takumi
T
T
Hiramoto
N
N
Iwasaka
T
T
Matsuda
H
H
D000200:Action Potentials / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D019556:COS Cells; D027682:Cation Transport Proteins; D004268:DNA-Binding Proteins; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D015870:Gene Expression / Q000502:physiology; D006352:Heart Ventricles / Q000166:cytology; D051656:KCNQ Potassium Channels; D051657:KCNQ1 Potassium Channel; D051379:Mice; D008813:Mice, Inbred ICR; D018485:Muscle Fibers, Skeletal / Q000502:physiology*; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D009206:Myocardium / Q000166:cytology*; D020319:Oligodeoxyribonucleotides, Antisense; D018408:Patch-Clamp Techniques; D010641:Phenotype; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism; D024642:Potassium Channels, Voltage-Gated; D015534:Trans-Activators; D014162:Transfection
D027682:Cation Transport Proteins; D004268:DNA-Binding Proteins; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; C000606913:KCNH2 protein, human; C092509:KCNH6 protein, human; D051656:KCNQ Potassium Channels; D051657:KCNQ1 Potassium Channel; C000606915:Kcnh2 protein, mouse; C496910:Kcnq1 protein, mouse; D020319:Oligodeoxyribonucleotides, Antisense; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D015534:Trans-Activators; C059983:potassium channel protein I(sk)