title: In calcineurin-induced cardiac hypertrophy expression of Nav1.5, Cx40 and Cx43 is reduced by different mechanisms.
abstract: Alterations in expression levels of Na(v)1.5, Cx43 and Cx40 have been frequently reported in cardiac disease and are associated with the development of arrhythmias, but little is known about the underlying molecular mechanisms. In this study we investigated electrical conduction and expression of Na(v)1.5, Cx43 and Cx40 in hearts of transgenic mice overexpressing a constitutively active form of calcineurin (MHC-CnA). ECG recordings showed that atrial, atrioventricular and ventricular activation were significantly prolonged in MHC-CnA hearts as compared to wildtype (WT) littermates. Epicardial activation and arrhythmia susceptibility analysis revealed increased ventricular activation thresholds and arrhythmia vulnerability. Moreover, epicardial ventricular activation patterns in MHC-CnA mice were highly discontinuous with multiple areas of block. These impaired conduction properties were associated with severe reductions in Na(v)1.5, Cx43 and Cx40 protein expression in MHC-CnA hearts as visualized by immunohistochemistry and immunoblotting. Real-time RT-PCR demonstrated that the decreased protein levels for Na(v)1.5 and Cx40, but not for Cx43, were accompanied by corresponding reductions at the RNA level. Cx43 RNA isoform analysis indicated that the reduction in Cx43 protein expression is caused by a post-transcriptional mechanism rather than by RNA isoform switching. In contrast, RNA isoform analysis for Cx40 and Na(v)1.5 provided additional evidence that in calcineurin-induced hypertrophy the downregulation of these proteins originates at the transcriptional level. These results provide the molecular rationale for Na(v)1.5, Cx43 and Cx40 downregulation in this model of hypertrophy and failure and the development of the pro-arrhythmic substrate.
authors.affiliation: Department of Medical Physiology, Division Heart & Lungs, University Medical Center Utrecht, Yalelaan 50, 3584 CM Utrecht, The Netherlands. M.F.A.Bierhuizen@umcutrecht.nl
authors.forename: Marti F A
authors.initials: MF
authors.lastname: Bierhuizen
authors.forename: Mohamed
authors.initials: M
authors.lastname: Boulaksil
authors.forename: Leonie
authors.initials: L
authors.lastname: van Stuijvenberg
authors.forename: Roel
authors.initials: R
authors.lastname: van der Nagel
authors.forename: Anita T
authors.initials: AT
authors.lastname: Jansen
authors.forename: Nancy A M
authors.initials: NA
authors.lastname: Mutsaers
authors.forename: Cansu
authors.initials: C
authors.lastname: Yildirim
authors.forename: Toon A B
authors.initials: TA
authors.lastname: van Veen
authors.forename: Leon J
authors.initials: LJ
authors.lastname: de Windt
authors.forename: Marc A
authors.initials: MA
authors.lastname: Vos
authors.forename: Harold V M
authors.initials: HV
authors.lastname: van Rijen
mesh_terms: D000818:Animals; D019703:Calcineurin / Q000009:adverse effects*; D006332:Cardiomegaly / Q000139:chemically induced / Q000378:metabolism*; D018031:Connexin 43 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics*; D017630:Connexins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics*; D015536:Down-Regulation / Q000502:physiology*; D005260:Female; D051379:Mice; D062554:NAV1.5 Voltage-Gated Sodium Channel; D015222:Sodium Channels / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic / Q000502:physiology; D000097003:Gap Junction alpha-5 Protein
chemical_list: D019703:Calcineurin; D018031:Connexin 43; D017630:Connexins; D062554:NAV1.5 Voltage-Gated Sodium Channel; D015222:Sodium Channels; D000097003:Gap Junction alpha-5 Protein; C568321:Scn5a protein, mouse