Phosphodiesterase 5 restricts NOS3/Soluble guanylate cyclase signaling to L-type Ca2+ current in cardiac myocytes

Honglan Wang, Mark J. Kohr, Christopher J. Traynham, Mark T. Ziolo

Research output: Contribution to journalArticlepeer-review

Abstract

Endothelial nitric oxide synthase (NOS3) regulates the functional response to β-adrenergic (β-AR) stimulation via modulation of the L-type Ca2+ current (ICa). However, the NOS3 signaling pathway modulating ICa is unknown. This study investigated the contribution of soluble guanylate cyclase (sGC) and phosphodiesterase type 5 (PDE5), a cGMP-specific PDE, in the NOS3-mediated regulation of ICa. Myocytes were isolated from NOS3 knockout (NOS3-/-) and wildtype (WT) mice. We measured ICa (whole-cell voltage-clamp), and simultaneously measured Ca2+ transients (Fluo-4 AM) and cell shortening (edge detection). Zaprinast (selective inhibitor of PDE5), decreased β-AR stimulated (isoproterenol, ISO)-ICa, and Ca2+ transient and cell shortening amplitudes in WT myocytes. However, YC-1 (NO-independent activator of sGC) only reduced ISO-stimulated ICa, but not cardiac contraction. We further investigated the NOS3/sGC/PDE5 pathway in NOS3-/- myocytes. PDE5 is mislocalized in these myocytes and we observed dissimilar effects of PDE5 inhibition and sGC activation compared to WT. That is, zaprinast had no effect on ISO-stimulated ICa, or Ca2+ transient and cell shortening amplitudes. Conversely, YC-1 significantly decreased both ISO-stimulated ICa, and cardiac contraction. Further confirming that PDE5 localizes NOS3/cGMP signaling to ICa; YC-1, in the presence of zaprinast, now significantly decreased ISO-stimulated Ca2+ transient and cell shortening amplitudes in WT myocytes. The effects of YC-1 on ICa and cardiac contraction were blocked by KT5823 (a selective inhibitor of the cGMP-dependent protein kinase, PKG). Our data suggests a novel physiological role for PDE5 in restricting the effects of NOS3/sGC/PKG signaling pathway to modulating β-AR stimulated ICa, while limiting effects on cardiac contraction.

Original languageEnglish (US)
Pages (from-to)304-314
Number of pages11
JournalJournal of Molecular and Cellular Cardiology
Volume47
Issue number2
DOIs
StatePublished - Aug 1 2009
Externally publishedYes

Keywords

  • Compartmentalization
  • Contraction
  • Cyclic GMP
  • L-type Ca channel
  • PDE5

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

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