Anti-minK antisense decreases the amplitude of the rapidly activating cardiac delayed rectifier K+ current.
The rapidly and slowly activating delayed rectifier K+ currents (IKr and IKs, respectively), which have different physiological properties have been identified in cardiac cells from several species, including humans. Although expression of the minimal K+ channel protein (minK) cDNA in some systems results in a current resembling IKs, the role of this gene product in channel function remains controversial. In atrial tumor myocytes (AT-1 cells), no IKs is recorded, but minK mRNA is detected, raising the possibility that expression of the minK gene serves an as-yet-unidentified function. In these experiments, AT-1 cells were exposed to antisense oligonucleotides targeting the 5' translation start site of the minK cDNA cloned from an AT-1 library. Cell size, IKr, and L-type and T-type Ca2+ currents were measured 24 to 48 hours after exposure and compared with data in cells exposed to the corresponding sense oligonucleotide or grown in medium only. Antisense oligonucleotide significantly reduced IKr compared with sense and medium-only control cells in 0 of 2 experiments (n = 3 to 6 cells per treatment in each experiment) at 50 nmol/L, 1 of 2 at 250 nmol/L, 6 of 6 at 1000 nmol/L, and 2 of 2 at 10,000 nmol/L. At 1000 nmol/L, maximum tail current in antisense-exposed cells was 2.5 +/- 0.1 pA/pF (mean +/- SEM, n = 28, 6 separate experiment), 6.6 +/- 0.4 pA/pF in sense-exposed cells (n = 27), 5.4 +/- 0.6 pA/pF in medium-only cells (n = 21), and 5.8 +/- 0.7 pA/pF in cells exposed to a random oligonucleotide (n = 9).(ABSTRACT TRUNCATED AT 250 WORDS)
Yang
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Vanderbilt University School of Medicine, Departments of Medicine and Pharmacology, Nashville, Tenn. 37232-6602, USA.
Kupershmidt
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Roden
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DM
D000818:Animals; D015220:Calcium Channels / Q000502:physiology; D002460:Cell Line / Q000166:cytology; D048429:Cell Size; D018076:DNA, Complementary / Q000235:genetics; D003627:Data Interpretation, Statistical; D004594:Electrophysiology; D005260:Female; D015870:Gene Expression; D006321:Heart / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism*; D016376:Oligonucleotides, Antisense; D016133:Polymerase Chain Reaction; D015221:Potassium Channels / Q000235:genetics*; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis
D015220:Calcium Channels; D018076:DNA, Complementary; D016376:Oligonucleotides, Antisense; D015221:Potassium Channels; D012333:RNA, Messenger; D012313:RNA