Amplified NO/cGMP-mediated relaxation and ryanodine receptor-to-BKCa channel signalling in corpus cavernosum smooth muscle from phospholamban knockout mice

Br J Pharmacol. 2012 Jan;165(2):455-66. doi: 10.1111/j.1476-5381.2011.01569.x.

Abstract

Background and purpose: Relaxation of corpus cavernosum smooth muscle (CCSM) is induced by NO. NO promotes the formation of cGMP, which activates cGMP-dependent protein kinase I (PKGI). The large conductance calcium-activated potassium (BK(Ca) ) channel is regarded as a major target of NO/cGMP signalling; however, the mechanism of BK(Ca) activation remains unclear. The aim of the present study was to determine whether sarcoplasmic reticulum (SR) Ca(2+) load and Ca(2+) release from the SR via ryanodine receptors (RyRs) is important for BK(Ca) channel activation in response to NO/cGMP.

Experimental approach: In vitro myography was performed on CCSM strips from wild-type and PLB knockout (PLB(-/-)) mice to evaluate contraction and relaxation in response to pharmacological agents and electrical field stimulation (EFS).

Key results: In CCSM strips from PLB(-/-) mice, a model of increased SR Ca(2+) load, contractile force in response to EFS or phenylephrine (PE) was increased by nearly 100%. EFS of strips precontracted with PE induced transient relaxation in CCSM, an effect that was significantly larger in PLB(-/-) strips. Likewise, the relaxation of PE-induced contraction in response to SNP and cGMP was greater in PLB(-/-) , as demonstrated by a shift in the concentration-response curve towards lower concentrations. Blocking RyRs and BK(Ca) channels diminished the induced relaxations and eliminated the difference between wild-type and PLB(-/-).

Conclusions and implications: NO/cGMP activates BK(Ca) channels through RyR-mediated Ca(2+) release. This signalling pathway is responsible for approximately 40% of the NO/cGMP effects and is amplified by increased SR Ca(2+) concentrations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenergic alpha-Agonists / pharmacology
  • Animals
  • Calcium / physiology
  • Calcium-Binding Proteins / deficiency
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / physiology
  • Cyclic GMP / physiology*
  • In Vitro Techniques
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / physiology*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Muscle Contraction / drug effects
  • Muscle Contraction / physiology*
  • Muscle, Smooth / drug effects
  • Muscle, Smooth / physiology*
  • Nitric Oxide / physiology*
  • Penis / drug effects
  • Penis / physiology
  • Phenylephrine / pharmacology
  • Ryanodine Receptor Calcium Release Channel / physiology*
  • Sarcoplasmic Reticulum / physiology
  • Signal Transduction

Substances

  • Adrenergic alpha-Agonists
  • BKCa protein, mouse
  • Calcium-Binding Proteins
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits
  • Ryanodine Receptor Calcium Release Channel
  • phospholamban
  • Phenylephrine
  • Nitric Oxide
  • Cyclic GMP
  • Calcium