Characterization of the role of HCN channels in β3-adrenoceptor mediated rat bladder relaxation
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Keywords
HCN, ZD7288, detrusor relaxation, β3-AR
Abstract
Objective: The second messenger cAMP is involved in both β3 adrenoceptor (β3-AR) mediated detrusor relaxation and the kinetics of Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels. Here we characterized the effect HCN channel activation and possible interaction with β3-AR in bladder.
Methods: Bladder tissues from Sprague-Dawley rats and Human organ donors obtained for studying species-specific expression of HCN channels by real-time QPCR and Western Blot. Effect of β3-agonist on rat bladder Strips (0.5 x 0.5 x 7 mm in size) was studied during activation and blockade of HCN channels by Lamotrigine and ZD7288, respectively.
Results: Expression of all four genes encoding for HCN channels (HCN1-4) was detected separately in bladder mucosa and detrusor from human and rat bladders. Species based differences shown by relatively higher expression of HCN4 isoform in human bladder and that of HCN1 in rat bladder. Western blot confirmed the findings at mRNA level. Cumulative application β3-AR agonist CL316,243 produced a concentration dependent decrease in resting tension of rat bladder strips expressed as integral of mechanical activity. Pre-incubation of HCN channel blocker ZD 7288 opposed the relaxant effect of CL316,243, whereas co-administration of lamotrigine with CL316,243 at equal molar concentrations caused an additive decrease in resting tension. Cumulative addition of ZD7288 and lamotrigine in absence of CL316,243 showed opposing effects on detrusor contractility
Conclusions: Species-specific differences noted in expression of HCN channels in bladder. Opposing effects ZD7288 and Lamotrigine in the action of β3-AR agonist demonstrate possible functional interaction of HCN channels and β3-AR in detrusor contractility.
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References
2. Tyagi P, Tyagi V, Yoshimura N, Chancellor M, Yamaguchi O (2009) Beta3-adrenoceptor agonists for the treatment of overactive bladder. Drugs Fut 34: 635.
3. Andersson KE, Arner A (2004) Urinary bladder contraction and relaxation: physiology and pathophysiology. Physiol Rev 84: 935-986.
4. Cuttle MF, Rusznak Z, Wong AY, Owens S, Forsythe ID (2001) Modulation of a presynaptic hyperpolarization-activated cationic current (I(h)) at an excitatory synaptic terminal in the rat auditory brainstem. J Physiol 534: 733-744.
5. Greenwood IA, Prestwich SA (2002) Characteristics of hyperpolarization-activated cation currents in portal vein smooth muscle cells. Am J Physiol Cell Physiol 282: C744-753.
6. Xue L, Li Y, Han X, Yao L, Yuan J, et al. (2012) Investigation of hyperpolarization-activated cyclic nucleotide-gated channels in interstitial cells of Cajal of human bladder. Urology 80: 224 e213-228.
7. Tyagi P, Thomas CA, Yoshimura N, Chancellor MB (2009) Investigations into the presence of functional Beta1, Beta2 and Beta3-adrenoceptors in urothelium and detrusor of human bladder. Int Braz J Urol 35: 76-83.
8. Poolos NP, Migliore M, Johnston D (2002) Pharmacological upregulation of h-channels reduces the excitability of pyramidal neuron dendrites. Nat Neurosci 5: 767-774.
9. Postea O, Biel M (2011) Exploring HCN channels as novel drug targets. Nat Rev Drug Discov 10: 903-914.
10. Surges R, Freiman TM, Feuerstein TJ (2003) Gabapentin increases the hyperpolarization-activated cation current Ih in rat CA1 pyramidal cells. Epilepsia 44: 150-156.
11. Tyagi V, Philips BJ, Su R, Smaldone MC, Erickson VL, et al. (2009) Differential expression of functional cannabinoid receptors in human bladder detrusor and urothelium. J Urol 181: 1932-1938.
12. Green ME, Edwards G, Kirkup AJ, Miller M, Weston AH (1996) Pharmacological characterization of the inwardly-rectifying current in the smooth muscle cells of the rat bladder. Br J Pharmacol 119: 1509-1518.
13. He P, Deng J, Zhong X, Zhou Z, Song B, et al. (2012) Identification of a hyperpolarization-activated cyclic nucleotide-gated channel and its subtypes in the urinary bladder of the rat. Urology 79: 1411 e1417-1413.
14. Xiao J, Nguyen TV, Ngui K, Strijbos PJ, Selmer IS, et al. (2004) Molecular and functional analysis of hyperpolarisation-activated nucleotide-gated (HCN) channels in the enteric nervous system. Neuroscience 129: 603-614.
15. Masuda N, Hayashi Y, Matsuyoshi H, Chancellor MB, de Groat WC, et al. (2006) Characterization of hyperpolarization-activated current (Ih) in dorsal root ganglion neurons innervating rat urinary bladder. Brain Res 1096: 40-52.
16. Gu N, Vervaeke K, Hu H, Storm JF (2005) Kv7/KCNQ/M and HCN/h, but not KCa2/SK channels, contribute to the somatic medium after-hyperpolarization and excitability control in CA1 hippocampal pyramidal cells. J Physiol 566: 689-715.
17. de Groat WC, Yoshimura N (2006) Mechanisms underlying the recovery of lower urinary tract function following spinal cord injury. Prog Brain Res 152: 59-84.
18. Sui G, Fry CH, Malone-Lee J, Wu C (2009) Aberrant Ca2+ oscillations in smooth muscle cells from overactive human bladders. Cell Calcium 45: 456-464.
19. Li Y, Xue L, Miao Q, Mao F, Yao L, et al. (2013) Expression and electrophysiological characteristics of P2X3 receptors in interstitial cells of Cajal in rats with partial bladder outlet obstruction. BJU Int 111: 843-851.
20. BoSmith RE, Briggs I, Sturgess NC (1993) Inhibitory actions of ZENECA ZD7288 on whole-cell hyperpolarization activated inward current (If) in guinea-pig dissociated sinoatrial node cells. Br J Pharmacol 110: 343-349.
21. Ghamari-Langroudi M, Bourque CW (2000) Excitatory role of the hyperpolarization-activated inward current in phasic and tonic firing of rat supraoptic neurons. J Neurosci 20: 4855-4863.
22. Deng T, Zhang Q, Wang Q, Zhong X, Li L (2015) Changes in hyperpolarization-activated cyclic nucleotide-gated channel expression and activity in bladder interstitial cells of Cajal from rats with detrusor overactivity. Int Urogynecol J.
23. Felix R, Sandoval A, Sanchez D, Gomora JC, De la Vega-Beltran JL, et al. (2003) ZD7288 inhibits low-threshold Ca(2+) channel activity and regulates sperm function. Biochem Biophys Res Commun 311: 187-192.
24. Albertson AJ, Yang J, Hablitz JJ (2011) Decreased hyperpolarization-activated currents in layer 5 pyramidal neurons enhances excitability in focal cortical dysplasia. J Neurophysiol 106: 2189-2200.
25. Kanai A, Roppolo J, Ikeda Y, Zabbarova I, Tai C, et al. (2007) Origin of spontaneous activity in neonatal and adult rat bladders and its enhancement by stretch and muscarinic agonists. Am J Physiol Renal Physiol 292: F1065-1072.
26. McCloskey KD, Gurney AM (2002) Kit positive cells in the guinea pig bladder. J Urol 168: 832-836.
27. Smet PJ, Jonavicius J, Marshall VR, de Vente J (1996) Distribution of nitric oxide synthase-immunoreactive nerves and identification of the cellular targets of nitric oxide in guinea-pig and human urinary bladder by cGMP immunohistochemistry. Neuroscience 71: 337-348.
28. Lentle RG, Reynolds GW, Janssen PW, Hulls CM, King QM, et al. (2015) Characterisation of the contractile dynamics of the resting ex vivo urinary bladder of the pig. BJU Int.
29. Zona C, Avoli M (1997) Lamotrigine reduces voltage-gated sodium currents in rat central neurons in culture. Epilepsia 38: 522-525.
30. Clouse AK, Jugus MJ, Eisennagel SH, Laping NJ, Westfall TD, et al. (2012) Voltage-gated Na+ channel blockers reduce functional bladder capacity in the conscious spontaneously hypertensive rat. Urology 79: 1410 e1411-1416.
31. Koh SD, Kim TW, Jun JY, Glasgow NJ, Ward SM, et al. (2000) Regulation of pacemaker currents in interstitial cells of Cajal from murine small intestine by cyclic nucleotides. J Physiol 527 Pt 1: 149-162.
32. Jun JY, Choi S, Yeum CH, Chang IY, Park CK, et al. (2004) Noradrenaline inhibits pacemaker currents through stimulation of beta 1-adrenoceptors in cultured interstitial cells of Cajal from murine small intestine. Br J Pharmacol 141: 670-677.
33. Dolan JA, Muenkel HA, Burns MG, Pellegrino SM, Fraser CM, et al. (1994) Beta-3 adrenoceptor selectivity of the dioxolane dicarboxylate phenethanolamines. J Pharmacol Exp Ther 269: 1000-1006.
34. Aizawa N, Igawa Y, Nishizawa O, Wyndaele JJ (2010) Effects of CL316,243, a beta 3-adrenoceptor agonist, and intravesical prostaglandin E2 on the primary bladder afferent activity of the rat. Neurourol Urodyn 29: 771-776.
35. Igawa Y, Aizawa N, Homma Y (2010) Beta3-adrenoceptor agonists: possible role in the treatment of overactive bladder. Korean J Urol 51: 811-818.
36. Loutochin O, Afraa TA, Campeau L, Mahfouz W, Elzayat E, et al. (2012) Effect of the anticonvulsant medications Pregabalin and Lamotrigine on Urodynamic parameters in an animal model of neurogenic detrusor overactivity. Neurourol Urodyn 31: 1197-1202.
37. Kuwabara Y, Kuwahara K, Takano M, Kinoshita H, Arai Y, et al. (2013) Increased expression of HCN channels in the ventricular myocardium contributes to enhanced arrhythmicity in mouse failing hearts. J Am Heart Assoc 2: e000150.
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Mahendra Kashyap, University of Pittsburgh
Post-doctoral ResearcherPhillip P. Smith, University of Connecticut Health Center, Farmington, CT,
Associate Professor, Department of Surgery,Michael Chancellor, William Beaumont School of Medicine
Professor, Department of UrologyPradeep Tyagi, University of Pittsburgh
Associate Professor,
Department of Urology, University of Pittsburgh
