Calcineurin increases cardiac transient outward K+ currents via transcriptional up-regulation of Kv4.2 channel subunits.
Fast transient outward potassium currents (I(to,f)) are critical determinants of regional heterogeneity of cardiomyocyte repolarization as well as cardiomyocyte contractility. Additionally, I(to,f) densities are markedly down-regulated in cardiac hypertrophy and heart disease, conditions associated with activation of the serine/threonine phosphatase calcineurin (Cn). In this study, we investigated the regulation of I(to,f) expression by Cn in cultured neonatal rat ventricular myocytes (NRVMs) with and without alpha(1)-adrenoreceptor stimulation with phenylephrine (PE). Overexpression of constitutively active Cn in NRVMs induced hypertrophy and caused profound increases in I(to,f) density as well as Kv4.2 mRNA and protein expression and promoter activity, without affecting Kv4.3 or KChIP2 levels. The effects of Cn on hypertrophy, I(to,f), and Kv4.2 transcription were associated with NFAT activation and were abrogated by NFAT inhibition. Despite activating Cn and inducing hypertrophy in NRVMs, PE resulted in profound down-regulation of I(to,f) densities as well as Kv4.2, Kv4.3, and KChIP2 expression. Although hypertrophy and NFAT activation were inhibited by the Cn inhibitory peptide CAIN, I(to,f) and Kv4.2 expression were further reduced by CAIN, whereas Cn overexpression eliminated PE-induced reductions in I(to,f) and Kv4.2 expression without affecting Kv4.3 or KChIP2 levels. We conclude that Cn increases cardiac I(to,f) densities by positively regulating Kv4.2 gene transcription. Consistent with this conclusion, we found that I(to,f) was increased in myocytes isolated from young mice overexpressing Cn prior to the development of heart disease. This positive regulation of Kv4.2 transcription by Cn activation is expected to minimize the reductions in I(to,f) and Kv4.2 expression observed in hypertrophic cardiomyocytes.
Gong
Nanling
N
Departments of Physiology and Medicine, Heart and Stroke/Richard Lewar Centre of Excellence, University Health Network, University of Toronto, 150 College Street, Toronto, Ontario M5S 3E2, Canada.
Bodi
Ilona
I
Zobel
Carsten
C
Schwartz
Arnold
A
Molkentin
Jeffery D
JD
Backx
Peter H
PH
D000818:Animals; D001483:Base Sequence; D019703:Calcineurin / Q000494:pharmacology*; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D009206:Myocardium / Q000378:metabolism*; D050778:NFATC Transcription Factors / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D051672:Shal Potassium Channels / Q000187:drug effects*; D014158:Transcription, Genetic; D015854:Up-Regulation
D017931:DNA Primers; D050778:NFATC Transcription Factors; D051672:Shal Potassium Channels; D019703:Calcineurin