β-adrenoceptor agonists stimulate endothelial nitric oxide synthase in rat urinary bladder urothelial cells

Lori A. Birder, Michele L. Nealen, Susanna Kiss, William C. De Groat, Michael Caterina, Edward Wang, Gerard Apodaca, Anthony J. Kanai

Research output: Contribution to journalArticle

Abstract

We have investigated the intracellular signaling mechanisms underlying the release of nitric oxide (NO) evoked by β-adrenoceptor (AR) agonists in urinary bladder strips and cultured bladder urothelial cells from adult rats. Reverse transcription-PCR revealed that inducible NO synthase and endothelial NOS but not neuronal NOS genes were expressed in urothelial cells. NO release from both urothelial cells and bladder strips was decreased (37-42%) in the absence of extracellular Ca2+ (100 μM EGTA) and was ablated after incubation with BAPTA-AM (5 μM) or caffeine (10 mM), indicating that the NO production is mediated in part by intracellular calcium stores. NO release was reduced (18-24%) by nifedipine (10 μM) and potentiated (29-32%) by incubation with the Ca2+ channel opener BAYK8644 (1-10 μM). In addition, β-AR-evoked NO release (isoproterenol; dobutamine; terbutaline; 10-9 to 10-5 M) was blocked by the NOS inhibitors NG-nitro-L-arginine methyl ester (30 μM) or NG-monomethyl-L-arginine (50 μM), by β-adrenoceptor antagonists (propranol, β12; atenolol, β1; ICl 118551; β2; 100 μM), or by the calmodulin antagonist trifluoperazine (50 μM). Incubating cells with the nonhydrolyzable GTP analog GTPγS (1 μM) or the membrane-permeant cAMP analog dibutyryl-cAMP (10-100 μM) directly evoked NO release. Forskolin (10 μM) or the phosphodiesterase IBMX (50 μM) enhanced (39-42%) agonist-evoked NO release. These results indicate that β-adrenoceptor stimulation activates the adenylate cyclase pathway in bladder epithelial cells and initiates an increase in intracellular Ca2+ that triggers NO production and release. These findings are considered in light of recent reports that urothelial cells may exhibit a number of "neuron-like" properties, including the expression of receptors/ion channels similar to those found in sensory neurons.

Original languageEnglish (US)
Pages (from-to)8063-8070
Number of pages8
JournalJournal of Neuroscience
Volume22
Issue number18
StatePublished - Sep 15 2002

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Adrenergic Receptors
Nitric Oxide
Urinary Bladder
omega-N-Methylarginine
Terbutaline
1-Methyl-3-isobutylxanthine
Trifluoperazine
Atenolol
Dobutamine
rat Nos3 protein
Egtazic Acid
NG-Nitroarginine Methyl Ester
Phosphoric Diester Hydrolases
Nitric Oxide Synthase Type II
Sensory Receptor Cells
Colforsin
Calmodulin
Nifedipine
Guanosine Triphosphate
Caffeine

Keywords

  • Adrenergic
  • ENOS
  • INOS
  • Nitric oxide
  • Urinary bladder
  • Urothelium

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Birder, L. A., Nealen, M. L., Kiss, S., De Groat, W. C., Caterina, M., Wang, E., ... Kanai, A. J. (2002). β-adrenoceptor agonists stimulate endothelial nitric oxide synthase in rat urinary bladder urothelial cells. Journal of Neuroscience, 22(18), 8063-8070.

β-adrenoceptor agonists stimulate endothelial nitric oxide synthase in rat urinary bladder urothelial cells. / Birder, Lori A.; Nealen, Michele L.; Kiss, Susanna; De Groat, William C.; Caterina, Michael; Wang, Edward; Apodaca, Gerard; Kanai, Anthony J.

In: Journal of Neuroscience, Vol. 22, No. 18, 15.09.2002, p. 8063-8070.

Research output: Contribution to journalArticle

Birder, LA, Nealen, ML, Kiss, S, De Groat, WC, Caterina, M, Wang, E, Apodaca, G & Kanai, AJ 2002, 'β-adrenoceptor agonists stimulate endothelial nitric oxide synthase in rat urinary bladder urothelial cells', Journal of Neuroscience, vol. 22, no. 18, pp. 8063-8070.
Birder, Lori A. ; Nealen, Michele L. ; Kiss, Susanna ; De Groat, William C. ; Caterina, Michael ; Wang, Edward ; Apodaca, Gerard ; Kanai, Anthony J. / β-adrenoceptor agonists stimulate endothelial nitric oxide synthase in rat urinary bladder urothelial cells. In: Journal of Neuroscience. 2002 ; Vol. 22, No. 18. pp. 8063-8070.
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