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  • Title: ATP- and adenosine-induced relaxation of the smooth muscle of the pig urethra.
    Author: Werkström V, Andersson KE.
    Journal: BJU Int; 2005 Dec; 96(9):1386-91. PubMed ID: 16287463.
    Abstract:
    OBJECTIVES: To investigate relaxation mechanisms for ATP and adenosine in the pig urethra, together with the possible role of ATP in nerve-evoked urethral relaxations, as ATP is thought to cause bladder smooth muscle contraction via P2X receptors, whereas relaxation is mediated via G-protein coupled P2Y receptors, and ATP may also induce relaxation via breakdown to adenosine. MATERIALS AND METHODS: Circular muscle strips from the female pig urethra were mounted in tissue baths to record force; the effects of increasing concentrations of 1-300 microM ATP, the P2-receptor agonist 2-methylthioATP (2-MeSATP), adenosine, the stable adenosine-analogue, 5'(N-ethylcarboxamido) adenosine (NECA), ADP, uridine-triphosphate (UTP) and alpha,beta-methylene-ATP were assessed on the spontaneously developed tone. Responses to ATP were further assessed in the presence of G-protein activator guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS; 1-10 microM), the G-protein inhibitor guanosine 5'-O-(2-thio-diphosphate) (GDPbetaS; 10-100 microM), suramin (1-100 microM), the ecto-ATPase inhibitor 6-N,N-diethyl-beta-gamma-dibromomethylene-D-adenosine-5-triphosphate (ARL 67156, 10-100 microM), and the suggested P2Y receptor antagonist, reactive blue-2 (1-100 microM). The effect of the adenosine (P1) receptor antagonist 8-(p-sulphophenyl)theophylline (8-SPT, 1-100 microM) on responses to adenosine, and the effects of the adenosine reuptake inhibitor S(p-nitrobenzyl)-6-thioinosine (NBTI, 1-100 microM) on responses to adenosine and ATP were also assessed. Responses to electrical field stimulation (EFS, 12 and 30 Hz) in the presence of phentolamine (1 microM), scopolamine (1 microM) and N omega-nitro-L-arginine (0.3 mM) were studied before and after treatment with GTPgammaS, GDPbetaS, suramin, reactive blue-2 and ARL 67156. RESULTS: Strips were relaxed in a concentration-dependent manner by exogenously administered ATP and 2-meSATP, the relaxations being slowly developing and long-lasting. The relaxant effect evoked by both agonists at 300 microM amounted to about half of the spontaneously developed tone. The relaxation evoked by ATP was not significantly affected by GTPgammaS, GDPbetaS, suramin, ARL 67156 or reactive blue-2. Adenosine induced a concentration-dependent relaxation of the smooth muscle tone, reaching a maximum of approximately 70% at 300 microM, whereas 300 microM NECA only relaxed the preparations by approximately 35%. The adenosine-induced relaxation was not affected by treatment with 8-SPT. However, NBTI (1 microM) significantly reduced the relaxation evoked by 300 microM adenosine. ADP relaxed the smooth muscle tone by approximately 40% (300 microM). There was no response to UTP, and the effect of alpha,beta-methylene-ATP was negligible (5% relaxation at 100 microM). EFS caused slowly developing and long-lasting relaxations that were unaffected by GTPgammaS, GDPbetaS, suramin, reactive blue-2 and ARL 67156. CONCLUSIONS: These results suggest that exogenous ATP and adenosine relax the smooth muscle of the pig urethra in a manner similar to that evoked by electrical stimulation of nerves, although there was no evidence for involvement of a definable P2Y receptor subtype in these relaxations.
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