title: Purinergic signalling underlies transforming growth factor-β-mediated bladder afferent nerve hyperexcitability.
abstract: The sensory components of the urinary bladder are responsible for the transduction of bladder filling and are often impaired with neurological injury or disease. Elevated extracellular ATP contributes, in part, to bladder afferent nerve hyperexcitability during urinary bladder inflammation or irritation. Transforming growth factor-β1 (TGF-β1) may stimulate ATP release from the urothelium through vesicular exocytosis mechanisms with minimal contribution from pannexin-1 channels to increase bladder afferent nerve discharge. Bladder afferent nerve hyperexcitability and urothelial ATP release with CYP-induced cystitis is decreased with TGF-β inhibition. These results establish a causal link between an inflammatory mediator, TGF-β, and intrinsic signalling mechanisms of the urothelium that may contribute to the altered sensory processing of bladder filling.
  
  
  
  The afferent limb of the micturition reflex is often compromised following bladder injury, disease and inflammatory conditions. We have previously demonstrated that transforming growth factor-β (TGF-β) signalling contributes to increased voiding frequency and decreased bladder capacity with cystitis. Despite the functional presence of TGF-β in bladder inflammation, the precise mechanisms of TGF-β mediating bladder dysfunction are not yet known. Thus, the present studies investigated the sensory components of the urinary bladder that may underlie the pathophysiology of aberrant TGF-β activation. We utilized bladder-pelvic nerve preparations to characterize bladder afferent nerve discharge and the mechanisms of urothelial ATP release with distention. Our findings indicate that bladder afferent nerve discharge is sensitive to elevated extracellular ATP during pathological conditions of urinary bladder inflammation or irritation. We determined that TGF-β1 may increase bladder afferent nerve excitability by stimulating ATP release from the urothelium via vesicular exocytosis mechanisms with minimal contribution from pannexin-1 channels. Furthermore, blocking aberrant TGF-β signalling in cyclophosphamide-induced cystitis with TβR-1 inhibition decreased afferent nerve hyperexcitability with a concomitant decrease in urothelial ATP release. Taken together, these results establish a role for purinergic signalling mechanisms in TGF-β-mediated bladder afferent nerve activation that may ultimately facilitate increased voiding frequency. The synergy between intrinsic urinary bladder signalling mechanisms and an inflammatory mediator provides novel insight into bladder dysfunction and supports new avenues for therapeutic intervention.
authors.affiliation: Department of Neurological Sciences, University of Vermont College of Medicine, Burlington, VT, 05405, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Gonzalez
authors.affiliation: Department of Pharmacology, University of Vermont College of Medicine, Burlington, VT, 05405, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Heppner
authors.affiliation: Department of Pharmacology, University of Vermont College of Medicine, Burlington, VT, 05405, USA.|Institute of Cardiovascular Sciences, University of Manchester, Manchester, UK.
authors.forename: Mark T
authors.initials: MT
authors.lastname: Nelson
authors.affiliation: Department of Neurological Sciences, University of Vermont College of Medicine, Burlington, VT, 05405, USA.
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Vizzard
mesh_terms: D000255:Adenosine Triphosphate / Q000502:physiology*; D000818:Animals; D017630:Connexins / Q000502:physiology; D003520:Cyclophosphamide; D003556:Cystitis / Q000139:chemically induced / Q000503:physiopathology*; D008297:Male; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D058914:Purinergic Antagonists / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000077293:Receptor, Transforming Growth Factor-beta Type I; D011983:Receptors, Purinergic / Q000502:physiology*; D018125:Receptors, Transforming Growth Factor beta / Q000502:physiology; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000502:physiology*; D001743:Urinary Bladder / Q000294:innervation* / Q000502:physiology*; D019459:Urothelium / Q000502:physiology
chemical_list: D017630:Connexins; D009419:Nerve Tissue Proteins; C503631:Panx1 protein, mouse; D058914:Purinergic Antagonists; D011983:Receptors, Purinergic; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D011732:Pyridoxal Phosphate; D000255:Adenosine Triphosphate; D003520:Cyclophosphamide; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I