NS309 decreases rat detrusor smooth muscle membrane potential and phasic contractions by activating SK3 channels

Br J Pharmacol. 2013 Apr;168(7):1611-25. doi: 10.1111/bph.12049.

Abstract

Background and purpose: Overactive bladder (OAB) is often associated with abnormally increased detrusor smooth muscle (DSM) contractions. We used NS309, a selective and potent opener of the small or intermediate conductance Ca(2+) -activated K(+) (SK or IK, respectively) channels, to evaluate how SK/IK channel activation modulates DSM function.

Experimental approach: We employed single-cell RT-PCR, immunocytochemistry, whole cell patch-clamp in freshly isolated rat DSM cells and isometric tension recordings of isolated DSM strips to explore how the pharmacological activation of SK/IK channels with NS309 modulates DSM function.

Key results: We detected SK3 but not SK1, SK2 or IK channels expression at both mRNA and protein levels by RT-PCR and immunocytochemistry in DSM single cells. NS309 (10 μM) significantly increased the whole cell SK currents and hyperpolarized DSM cell resting membrane potential. The NS309 hyperpolarizing effect was blocked by apamin, a selective SK channel inhibitor. NS309 inhibited the spontaneous phasic contraction amplitude, force, frequency, duration and tone of isolated DSM strips in a concentration-dependent manner. The inhibitory effect of NS309 on spontaneous phasic contractions was blocked by apamin but not by TRAM-34, indicating no functional role of the IK channels in rat DSM. NS309 also significantly inhibited the pharmacologically and electrical field stimulation-induced DSM contractions.

Conclusions and implications: Our data reveal that SK3 channel is the main SK/IK subtype in rat DSM. Pharmacological activation of SK3 channels with NS309 decreases rat DSM cell excitability and contractility, suggesting that SK3 channels might be potential therapeutic targets to control OAB associated with detrusor overactivity.

Publication types

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

MeSH terms

  • Animals
  • Apamin / pharmacology
  • Cell Size / drug effects
  • Electric Stimulation
  • Female
  • In Vitro Techniques
  • Indoles / pharmacology*
  • Intermediate-Conductance Calcium-Activated Potassium Channels / antagonists & inhibitors
  • Intermediate-Conductance Calcium-Activated Potassium Channels / metabolism*
  • Ion Channel Gating
  • Male
  • Membrane Potentials / drug effects
  • Muscle Contraction / drug effects
  • Muscle, Smooth / drug effects*
  • Muscle, Smooth / physiology
  • Myocytes, Smooth Muscle / cytology
  • Myocytes, Smooth Muscle / drug effects
  • Myocytes, Smooth Muscle / physiology
  • Oximes / pharmacology*
  • Pyrazoles / pharmacology
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Small-Conductance Calcium-Activated Potassium Channels / antagonists & inhibitors
  • Small-Conductance Calcium-Activated Potassium Channels / genetics
  • Small-Conductance Calcium-Activated Potassium Channels / metabolism*
  • Urinary Bladder / drug effects*
  • Urinary Bladder / physiology

Substances

  • 6,7-dichloro-1H-indole-2,3-dione 3-oxime
  • Indoles
  • Intermediate-Conductance Calcium-Activated Potassium Channels
  • Kcnn3 protein, rat
  • Oximes
  • Pyrazoles
  • RNA, Messenger
  • Small-Conductance Calcium-Activated Potassium Channels
  • TRAM 34
  • Apamin