View More View Less
  • 1 Paracelsus Medical University, Austria
  • | 2 University Hospital Muenster, Germany
  • | 3 University Clinic of Ophthalmology and Optometry, Paracelsus Medical University/SALK, Austria
  • | 4 Bashkir State Medical University, Russia
Restricted access

Purchase article

USD  $25.00

1 year subscription (Individual Only)

USD  $752.00

Background

Caveolin-3 (cav-3) mutations are linked to the long-QT syndrome (LQTS) causing distinct clinical symptoms. Hyperpolarization-activated cyclic nucleotide channel 4 (HCN4) underlies the pacemaker current If. It associates with cav-3 and both form a macromolecular complex.

Methods

To examine the effects of human LQTS-associated cav-3 mutations on HCN4-channel function, HEK293-cells were cotransfected with HCN4 and wild-type (WT) cav-3 or a LQTS-associated cav-3 mutant (T78M, A85T, S141R, or F97C). HCN4 currents were recorded using the whole-cell patch-clamp technique.

Results

WT cav-3 significantly decreased HCN4 current density and shifted midpoint of activation into negative direction. HCN4 current properties were differentially modulated by LQTS-associated cav-3 mutations. When compared with WT cav-3, A85T, F97C, and T78M did not alter the specific effect of cav-3, but S141R significantly increased HCN4 current density. Compared with WT cav-3, no significant modifications of voltage dependence of steady-state activation curves were observed. However, while WT cav-3 alone had no significant effect on HCN4 current activation, all LQTS-associated cav-3 mutations significantly accelerated HCN4 activation kinetics.

Conclusions

Our results indicate that HCN4 channel function is modulated by cav-3. LQTS-associated mutations of cav-3 differentially influence pacemaker current properties indicating a pathophysiological role in clinical manifestations.

  • 1.

    Abbott GW , Sesti F , Splawski I , Buck ME , Lehmann MH , Timothy KW , Keating MT , Goldstein SA : MiRP1 forms IKr potassium channels with HERG and is associated with cardiac arrhythmia. Cell 97, 175187 (1999)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Anderson RG : The caveolae membrane system. Annu. Rev. Biochem. 67, 199225 (1998)

  • 3.

    Balijepalli RC , Foell JD , Hall DD , Hell JW , Kamp TJ : Localization of cardiac L-type Ca2+ channels to a caveolar macromolecular signaling complex is required for β2-adrenergic regulation. Proc. Natl. Acad. Sci. U S A 103, 75007505 (2006)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4.

    Barbuti A , Gravante B , Riolfo M , Milanesi R , Terragni B , DiFrancesco D : Localization of pacemaker channels in lipid rafts regulates channel kinetics. Circ. Res. 94, 13251331 (2004)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 5.

    Barbuti A , Terragni B , Brioschi C , DiFrancesco D : Localization of f-channels to caveolae mediates specific β2-adrenergic receptor modulation of rate in sinoatrial myocytes. J. Mol. Cell. Cardiol. 42, 7178 (2007)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Bosman A , Sartiani L , Spinelli V , Del Lungo M , Stillitano F , Nosi D , Mugelli A , Cerbai E , Jaconi M : Molecular and functional evidence of HCN4 and caveolin-3 interaction during cardiomyocyte differentiation from human embryonic stem cells. Stem Cells Dev. 22, 17171727 (2013)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7.

    Brandt MC , Endres-Becker J , Zagidullin N , Motloch LJ , Er F , Rottlaender D , Michels G , Herzig S , Hoppe UC : Effects of KCNE2 on HCN isoforms: distinct modulation of membrane expression and single channel properties. Am. J. Physiol. Heart Circ. Physiol. 297, H355H363 (2009)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8.

    Er F , Larbig R , Ludwig A , Biel M , Hofmann F , Beuckelmann DJ , Hoppe UC : Dominant-negative suppression of HCN channels markedly reduces the native pacemaker current If and undermines spontaneous beating of neonatal cardiomyocytes. Circulation 107, 485489 (2003)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Galbiati F , Engelman JA , Volonte D , Zhang XL , Minetti C , Li M , Hou H Jr , Kneitz B , Edelmann W , Lisanti MP : Caveolin-3 null mice show a loss of caveolae, changes in the microdomain distribution of the dystrophin-glycoprotein complex, and t-tubule abnormalities. J. Biol. Chem. 276, 2142521433 (2001)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Gao LL , McMullan S , Djouhri L , Acosta C , Harper AA , Lawson SN : Expression and properties of hyperpolarization-activated current in rat dorsal root ganglion neurons with known sensory function. J. Physiol. 590, 46914705 (2012)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Greener A , Callahan M , Jerpseth B : An efficient random mutagenesis technique using an E. coli mutator strain. Mol. Biotechnol. 7, 189195 (1997)

  • 12.

    Harder T , Simons K : Caveolae, DIGs, and the dynamics of sphingolipid-cholesterol microdomains. Curr. Opin. Cell Biol. 9, 534542 (1997)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Hnasko R , Lisanti MP : The biology of caveolae: lessons from caveolin knockout mice and implications for human disease. Mol. Interv. 3, 445464 (2003)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14.

    Hoppe UC , Jansen E , Sudkamp M , Beuckelmann DJ : Hyperpolarization-activated inward current in ventricular myocytes from normal and failing human hearts. Circulation 97, 5565 (1998)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15.

    Hoppe UC , Marban E , Johns DC : Distinct gene-specific mechanisms of arrhythmia revealed by cardiac gene transfer of two long QT disease genes, HERG and KCNE1. Proc. Natl. Acad. Sci. U S A 98, 53355340 (2001)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Isbrandt D , Friederich P , Solth A , Haverkamp W , Ebneth A , Borggrefe M , Funke H , Sauter K , Breithardt G , Pongs O , Schulze-Bahr E : Identification and functional characterization of a novel KCNE2 (MiRP1) mutation that alters HERG channel kinetics. J. Mol. Med. 80, 524532 (2002)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Kuwabara Y , Kuwahara K , Takano M , Kinoshita H , Arai Y , Yasuno S , Nakagawa Y , Igata S , Usami S , Minami T , Yamada Y , Nakao K , Yamada C , Shibata J , Nishikimi T , Ueshima K , Nakao K : Increased expression of HCN channels in the ventricular myocardium contributes to enhanced arrhythmicity in mouse failing hearts. J. Am. Heart Assoc. 2, e000150 (2013)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Ludwig A , Zong X , Stieber J , Hullin R , Hofmann F , Biel M : Two pacemaker channels from human heart with profoundly different activation kinetics. EMBO J. 18, 23232329 (1999)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19.

    Martens JR , Sakamoto N , Sullivan SA , Grobaski TD , Tamkun MM : Isoform-specific localization of voltage-gated K+ channels to distinct lipid raft populations. Targeting of Kv1.5 to caveolae. J. Biol. Chem. 276, 84098414 (2001)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 20.

    Masson-Pevet M , Gros D , Besselsen E : The caveolae in rabbit sinus node and atrium. Cell Tissue Res. 208, 183196 (1980)

  • 21.

    Michels G , Er F , Khan IF , Endres-Becker J , Brandt MC , Gassanov N , Johns DC , Hoppe UC : K+ channel regulator KCR1 suppresses heart rhythm by modulating the pacemaker current If. PLoS One 3, e1511 (2008)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 22.

    Mitrofanova LB , Gorshkov AN , Lebedev DS , Mikhaylov EN : Evidence of specialized tissue in human interatrial septum: histological, immunohistochemical and ultrastructural findings. PLoS One 9, e113343 (2014)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23.

    Plotnikov AN , Bucchi A , Shlapakova I , Danilo P Jr , Brink PR , Robinson RB , Cohen IS , Rosen MR : HCN212-channel biological pacemakers manifesting ventricular tachyarrhythmias are responsive to treatment with If blockade. Heart Rhythm 5, 282288 (2008)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24.

    Roden DM , Lazzara R , Rosen M , Schwartz PJ , Towbin J , Vincent GM : Multiple mechanisms in the long-QT syndrome. Current knowledge, gaps, and future directions. The SADS Foundation Task Force on LQTS. Circulation 94, 19962012 (1996)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Rybin VO , Xu X , Lisanti MP , Steinberg SF : Differential targeting of β-adrenergic receptor subtypes and adenylyl cyclase to cardiomyocyte caveolae. A mechanism to functionally regulate the cAMP signaling pathway. J. Biol. Chem. 275, 4144741457 (2000)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Shi W , Wymore R , Yu H , Wu J , Wymore RT , Pan Z , Robinson RB , Dixon JE , McKinnon D , Cohen IS : Distribution and prevalence of hyperpolarization-activated cation channel (HCN) mRNA expression in cardiac tissues. Circ. Res. 85, e1e6 (1999)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27.

    Task Force for the Diagnosis and Management of Syncope, European Society of Cardiology (ESC), European Heart Rhythm Association (EHRA), Heart Failure Association (HFA), Heart Rhythm Society (HRS), Moya A, Sutton R, Ammirati F, Blanc JJ, Brignole M, Dahm JB, Deharo JC, Gajek J, Gjesdal K, Krahn A, Massin M, Pepi M, Pezawas T, Ruiz Granell R, Sarasin F, Ungar A, van Dijk JG, Walma EP, Wieling W: Guidelines for the diagnosis and management of syncope (version 2009). Eur. Heart J. 30, 26312671 (2009)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    Towbin JA , Vatta M : Molecular biology and the prolonged QT syndromes. Am. J. Med. 110, 385398 (2001)

  • 29.

    Vatta M , Ackerman MJ , Ye B , Makielski JC , Ughanze EE , Taylor EW , Tester DJ , Balijepalli RC , Foell JD , Li Z , Kamp TJ , Towbin JA : Mutant caveolin-3 induces persistent late sodium current and is associated with long-QT syndrome. Circulation 114, 21042112 (2006)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30.

    Yarbrough TL , Lu T , Lee HC , Shibata EF : Localization of cardiac sodium channels in caveolin-rich membrane domains: regulation of sodium current amplitude. Circ. Res. 90, 443449 (2002)

    • Crossref
    • Search Google Scholar
    • Export Citation
All Time Past Year Past 30 Days
Abstract Views 75 75 13
Full Text Views 22 9 0
PDF Downloads 20 8 0