Sensitization of pelvic afferent nerves in the in vitro rat urinary bladder-pelvic nerve preparation by purinergic agonists and cyclophosphamide pretreatment.

Effects of purinergic agonists (alpha,beta-meATP and ATP) and cyclophosphamide-induced cystitis on bladder afferent nerve (BAN) activity were studied in an in vitro bladder-pelvic nerve preparation. Distension of the bladder induced spontaneous bladder contractions that were accompanied by multiunit afferent firing. Intravesical administration of 40 and 130 microM alpha,beta-meATP increased afferent firing from 27 +/- 3 to 53 +/- 6 and 61 +/- 2 spikes/s, respectively, but did not change the maximum amplitude of spontaneous bladder contractions. Electrical stimulation on the surface of the bladder elicited action potentials (AP) in BAN. alpha,beta-meATP decreased the voltage threshold from 9.0 +/- 1.2 to 3.5 +/- 0.5 V (0.15-ms pulse duration) and increased the area of the APs (82% at 80-V stimulus intensity). These effects were blocked by TNP-ATP (30 microM). ATP (2 mM) applied in the bath produced similar changes in BAN activity. These effects were blocked by bath application of PPADS (30 microM). Neither TNP-ATP nor PPADS affected BAN activity induced by distension of the bladder. Cystitis induced by pretreatment of the rats with cyclophosphamide (100 mg/kg ip) increased afferent firing in response to isotonic bladder distension (10-40 cmH(2)O), decreased the threshold, and increased the area of evoked APs. The increase in afferent firing at 10 cmH(2)O intravesical pressure was reduced 52% by PPADS. These results indicate that purinergic agonists acting on P2X receptors and cystitis induced by cyclophosphamide can increase excitability of the BANs.

[1]  Ferguson.  Urothelial function , 1999, BJU international.

[2]  R. Burgstahler,et al.  ATP affects both axons and Schwann cells of unmyelinated C fibres , 2001, PAIN®.

[3]  P. Illés,et al.  Subsensitivity of P2X but not vanilloid 1 receptors in dorsal root ganglia of rats caused by cyclophosphamide cystitis. , 2003, European journal of pharmacology.

[4]  M. Vizzard.  Changes in Urinary Bladder Neurotrophic Factor mRNA and NGF Protein Following Urinary Bladder Dysfunction , 2000, Experimental Neurology.

[5]  G. Burnstock,et al.  Distribution of P2X receptors in the urinary bladder and the ureter of the rat. , 2000, The Journal of urology.

[6]  S. Brookes,et al.  Properties of the major classes of mechanoreceptors in the guinea pig bladder , 2007, The Journal of physiology.

[7]  W. D. de Groat,et al.  Increased expression of neuronal nitric oxide synthase in bladder afferent pathways following chronic bladder irritation , 1996, The Journal of comparative neurology.

[8]  G. Burnstock,et al.  Alterations in P2X and P2Y purinergic receptor expression in urinary bladder from normal cats and cats with interstitial cystitis. , 2004, American journal of physiology. Renal physiology.

[9]  Geoffrey Burnstock,et al.  Historical review: ATP as a neurotransmitter. , 2006, Trends in pharmacological sciences.

[10]  M. Koltzenburg,et al.  Activation of unmyelinated afferent fibres by mechanical stimuli and inflammation of the urinary bladder in the cat. , 1990, The Journal of physiology.

[11]  G. Somogyi,et al.  Enhanced ATP release from rat bladder urothelium during chronic bladder inflammation: Effect of botulinum toxin A , 2005, Neurochemistry International.

[12]  W. C. Groat.  A neurologic basis for the overactive bladder , 1997 .

[13]  S. Palea,et al.  Evidence for the presence of both pre‐ and postjunctional P2‐purinoceptor subtypes in human isolated urinary bladder , 1995, British journal of pharmacology.

[14]  J. Morrison.  The activation of bladder wall afferent nerves , 1999, Experimental physiology.

[15]  W. C. Groat,et al.  Effect of intravesical nitric oxide therapy on cyclophosphamide-induced cystitis. , 1999, The Journal of urology.

[16]  W. C. Groat.  The urothelium in overactive bladder: Passive bystander or active participant? , 2004 .

[17]  G. Burnstock,et al.  Activation and sensitisation of low and high threshold afferent fibres mediated by P2X receptors in the mouse urinary bladder , 2002, The Journal of physiology.

[18]  G. Burnstock,et al.  ATP is released from guinea pig ureter epithelium on distension. , 2002, American journal of physiology. Renal physiology.

[19]  J. Gosling,et al.  Sensory nerves in the mammalian urinary tract. An evaluation using light and electron microscopy. , 1974, Journal of anatomy.

[20]  W. D. de Groat,et al.  A neurologic basis for the overactive bladder. , 1997, Urology.

[21]  Yan Sun,et al.  Augmented extracellular ATP signaling in bladder urothelial cells from patients with interstitial cystitis. , 2006, American journal of physiology. Cell physiology.

[22]  K. Moore,et al.  Distribution and colocalization of calcitonin gene-related peptide, tachykinins, and vasoactive intestinal peptide in normal and idiopathic unstable human urinary bladder. , 1997, Laboratory investigation; a journal of technical methods and pathology.

[23]  I. Kennedy,et al.  ATP is released from rabbit urinary bladder epithelial cells by hydrostatic pressure changes–possible sensory mechanism? , 1997, The Journal of physiology.

[24]  G Burnstock,et al.  P2X receptors in sensory neurones. , 2000, British journal of anaesthesia.

[25]  W. D. de Groat.  The urothelium in overactive bladder: passive bystander or active participant? , 2004, Urology.

[26]  R. Pandita,et al.  Intravesical adenosine triphosphate stimulates the micturition reflex in awake, freely moving rats. , 2002, The Journal of urology.

[27]  G. Gabella,et al.  Distribution of afferent axons in the bladder of rats , 1998, Journal of neurocytology.

[28]  D. Cockayne,et al.  P2X3 Knock-Out Mice Reveal a Major Sensory Role for Urothelially Released ATP , 2001, The Journal of Neuroscience.

[29]  P. M. Lang,et al.  Functional Evidence for P2X Receptors in Isolated Human Vagus Nerve , 2003, Anesthesiology.

[30]  Namasivayam,et al.  Purinergic sensory neurotransmission in the urinary bladder: an in vitro study in the rat , 1999, BJU international.

[31]  A. Ahluwalia,et al.  Characterization of the capsaicin‐sensitive component of cyclophosphamide‐induced inflammation in the rat urinary bladder , 1994, British journal of pharmacology.

[32]  T. Maeda,et al.  Substance P-containing axon terminals in the mucosa of the human urinary bladder: pre-embedding immunohistochemistry using cryostat sections for electron microscopy , 1993, Histochemistry.

[33]  C. Maggi,et al.  Nervous control of the urogenital system , 1993 .

[34]  G. Burnstock.  Purinergic Signalling in Lower Urinary Tract , 2001 .

[35]  P. Cox.  Cyclophosphamide cystitis--identification of acrolein as the causative agent. , 1979, Biochemical pharmacology.

[36]  Robert Elde,et al.  Distinct ATP receptors on pain-sensing and stretch-sensing neurons , 1997, Nature.

[37]  J. Borlak,et al.  The myosin ATPase inhibitor 2,3-butanedione monoxime dictates transcriptional activation of ion channels and Ca(2+)-handling proteins. , 2004, Molecular pharmacology.

[38]  P. Reeh,et al.  Noxious heat‐induced CGRP release from rat sciatic nerve axons in vitro , 2001, The European journal of neuroscience.

[39]  Geoffrey Burnstock,et al.  Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X 3-deficient mice , 2000, Nature.

[40]  W. D. de Groat,et al.  Detrusor overactivity induced by intravesical application of adenosine 5'-triphosphate under different delivery conditions in rats. , 2005, Urology.

[41]  G. Lewin,et al.  Selective activation of nociceptors by P2X receptor agonists in normal and inflamed rat skin , 2001, The Journal of physiology.

[42]  D. Menétrey,et al.  Cyclophosphamide cystitis as a model of visceral pain in rats: model elaboration and spinal structures involved as revealed by the expression of c-Fos and Krox-24 proteins , 2004, Experimental Brain Research.

[43]  鈴木 秀佳.  Subtypes of purinoceptors in rat and dog urinary bladder smooth muscles , 1994 .

[44]  J. Roppolo,et al.  Feline interstitial cystitis results in mechanical hypersensitivity and altered ATP release from bladder urothelium. , 2003, American journal of physiology. Renal physiology.

[45]  S. McMahon,et al.  ATP in human skin elicits a dose-related pain response which is potentiated under conditions of hyperalgesia. , 2000, Brain : a journal of neurology.

[46]  S. Brookes,et al.  Major classes of sensory neurons to the urinary bladder , 2006, Autonomic Neuroscience.

[47]  A novel type of unmyelinated chemosensitive nociceptor in the acutely inflamed urinary bladder , 1988, Agents and Actions.

[48]  G. Gebhart,et al.  Mechanosensitive properties of pelvic nerve afferent fibers innervating the urinary bladder of the rat. , 1994, Journal of neurophysiology.

[49]  G. Burnstock,et al.  A unifying purinergic hypothesis for the initiation of pain , 1996, The Lancet.

[50]  P. Illés,et al.  P2X receptors and nociception. , 2001, Pharmacological reviews.

[51]  G. Burnstock,et al.  A P2X purinoceptor expressed by a subset of sensory neurons , 1995, Nature.