Pannexin 1 channels mediate the release of ATP into the lumen of the rat urinary bladder. (11th February 2015)

• Record Type: Journal Article
• Authors: Beckel, Jonathan M.; Daugherty, Stephanie L.; Tyagi, Pradeep; Wolf‐Johnston, Amanda S.; Birder, Lori A.; Mitchell, Claire H.; de Groat, William C.
• Is Part Of: Journal of physiology. Volume 593:Number 8(2015:Apr.)
• Page Start: 1857
• Page End: 1871
• Publication Date: 2015-02-11
• Subjects: Physiology -- Periodicals; 612.005
• DOI: 10.1113/jphysiol.2014.283119
• Languages: English
• ISSNs: 0022-3751

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="tjp6548-sec-0010" sec-type="section"> <title>Key points</title> <p> <list id="tjp6548-list-0001" list-type="bullet"> <list-item> <p>ATP is released through pannexin channels into the lumen of the rat urinary bladder in response to distension or stimulation with bacterial endotoxins.</p> </list-item> <list-item> <p>Luminal ATP plays a physiological role in the control of micturition because intravesical perfusion of apyrase or the ecto‐ATPase inhibitor ARL67156 altered reflex bladder activity in the anaesthetized rat.</p> </list-item> <list-item> <p>The release of ATP from the apical and basolateral surfaces of the urothelium appears to be mediated by separate mechanisms because intravesical administration of the pannexin channel antagonist Brilliant Blue FCF increased bladder capacity, whereas <sc>i.v.</sc> administration did not.</p> </list-item> <list-item> <p>Intravesical instillation of small interfering RNA‐containing liposomes decreased pannexin 1 expression in the rat urothelium <italic>in vivo</italic> and increased bladder capacity.</p> </list-item> <list-item> <p>These data indicate a role for pannexin‐mediated luminal ATP release in both the physiological and pathophysiological control of micturition and suggest that urothelial pannexin may be a viable target for the treatment of overactive bladder disorders.</p> </list-item> </list> </p> </sec> <sec id="tjp6548-sec-0020" sec-type="section"> <title>Abstract</title> <p>ATP is released from the bladder epithelium, also termed the urothelium, in response to mechanical or chemical stimuli. Although numerous studies have described the contribution of this release to the development of various bladder disorders, little information exists regarding the mechanisms of release. In the present study, we examined the role of pannexin channels in mechanically‐induced ATP release from the urothelium. PCR confirmed the presence of pannexin 1 and 2 mRNA in rat urothelial tissue, whereas immunofluorescence experiments localized pannexin 1 to all three layers of the urothelium. During continuous bladder cystometry in anaesthetized rats, inhibition of pannexin 1 channels using carbenoxolone (CBX) or Brilliant Blue FCF (BB‐FCF) (1–100 μ<sc>m</sc>, intravesically), or by using intravesical small interfering RNA, increased the interval between voiding contractions. Intravenous administration of BB‐FCF (1–100 μg kg<sup>−1</sup>) did not alter bladder activity. CBX or BB‐FCF (100 μ<sc>m</sc> intravesically) also decreased basal ATP concentrations in the perfusate from non‐distended bladders and inhibited increases in ATP concentrations in response to bladder distension (15 and 30 cmH<sub>2</sub>O pressure). Intravesical perfusion of the ATP diphosphohydrolase apyrase (2 U ml<sup>−1</sup>), or the ATPase inhibitor ARL67156 (10 μ<sc>m</sc>) increased or decreased reflex bladder activity, respectively. Intravesical instillation of bacterial lipopolysaccharides (LPS) (<italic>Escherichia coli</italic> 055:B5, 100 μg ml<sup>−1</sup>) increased ATP concentrations in the bladder perfusate, and also increased voiding frequency; these effects were suppressed by BB‐FCF. These data indicate that pannexin channels contribute to distension‐ or LPS‐evoked ATP release into the lumen of the bladder and that luminal release can modulate voiding function.</p> </sec> </abstract>