title: Expression and function of K(V)2-containing channels in human urinary bladder smooth muscle.
abstract: The functional role of the voltage-gated K(+) (K(V)) channels in human detrusor smooth muscle (DSM) is largely unexplored. Here, we provide molecular, electrophysiological, and functional evidence for the expression of K(V)2.1, K(V)2.2, and the electrically silent K(V)9.3 subunits in human DSM. Stromatoxin-1 (ScTx1), a selective inhibitor of K(V)2.1, K(V)2.2, and K(V)4.2 homotetrameric channels and of K(V)2.1/9.3 heterotetrameric channels, was used to examine the role of these channels in human DSM function. Human DSM tissues were obtained during open bladder surgeries from patients without a history of overactive bladder. Freshly isolated human DSM cells were studied using RT-PCR, immunocytochemistry, live-cell Ca(2+) imaging, and the perforated whole cell patch-clamp technique. Isometric DSM tension recordings of human DSM isolated strips were conducted using tissue baths. RT-PCR experiments showed mRNA expression of K(V)2.1, K(V)2.2, and K(V)9.3 (but not K(V)4.2) channel subunits in human isolated DSM cells. K(V)2.1 and K(V)2.2 protein expression was confirmed by Western blot analysis and immunocytochemistry. Perforated whole cell patch-clamp experiments revealed that ScTx1 (100 nM) inhibited the amplitude of the voltage step-induced K(V) current in freshly isolated human DSM cells. ScTx1 (100 nM) significantly increased the intracellular Ca(2+) level in DSM cells. In human DSM isolated strips, ScTx1 (100 nM) increased the spontaneous phasic contraction amplitude and muscle force, and enhanced the amplitude of the electrical field stimulation-induced contractions within the range of 3.5-30 Hz stimulation frequencies. These findings reveal that ScTx1-sensitive K(V)2-containing channels are key regulators of human DSM excitability and contractility and may represent new targets for pharmacological or genetic intervention for bladder dysfunction.
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, Columbia, 29208, USA.
authors.forename: Kiril L
authors.initials: KL
authors.lastname: Hristov
authors.forename: Muyan
authors.initials: M
authors.lastname: Chen
authors.forename: Serge A Y
authors.initials: SA
authors.lastname: Afeli
authors.forename: Qiuping
authors.initials: Q
authors.lastname: Cheng
authors.forename: Eric S
authors.initials: ES
authors.lastname: Rovner
authors.forename: Georgi V
authors.initials: GV
authors.lastname: Petkov
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D004558:Electric Stimulation; D005260:Female; D006801:Humans; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008875:Middle Aged; D009119:Muscle Contraction / Q000502:physiology; D009130:Muscle, Smooth / Q000378:metabolism / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000379:methods; D010455:Peptides; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051669:Shab Potassium Channels / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D013111:Spider Venoms; D001743:Urinary Bladder / Q000378:metabolism / Q000502:physiology*; D055815:Young Adult
chemical_list: D010455:Peptides; D012333:RNA, Messenger; D051669:Shab Potassium Channels; D013111:Spider Venoms; C560803:stromatoxin protein, Stromatopelma calceata