Demonstration of 5-HT(3) receptor function and expression in the mouse bladder.
The aim of this study was to demonstrate the presence of 5-HT(3) receptors in the mouse bladder and to determine their location. Bladder strips from female mice were set up in gassed Krebs-Henseleit solution at 37 degrees C and contractions recorded in response to electrical field stimulation (8 Hz, 60 V, 0.5-ms pulse duration) applied for 2 s every 50 s. The potentiating effects of 5-hydroxytryptamine (5-HT) were recorded (in the presence of 1-microM methysergide and 1-microM GR125487 to isolate the 5-HT(3) receptor response), and contractions were expressed as a percentage of the response to 0.1-M KCl. Responses to (5-HT) were also obtained in the presence of the 5-HT(3) receptor antagonist, ondansetron. RT-PCR was used to detect the expression of the 5-HT(3A) and 5-HT(3B) subunit transcripts of the mouse 5-HT(3) receptor. 5-HT and 5-HT(3) receptor agonists caused concentration-dependent increases in the force of neurogenic contractions without affecting the baseline tone. The rank order of potency was: meta-chloro-phenylbiguanide (m-CPB) = 5-HT > 2-methyl-5-HT (2m5-HT) = 1-phenylbiguanide (1-PBG). The respective pEC(50) values were: 6.42 +/- 0.2 = 5.95 +/- 0.19 > 5.35 +/- 0.12 = 5.14 +/- 0.13. m-CPB acted as a full agonist (E (max) = 40.65 +/- 3.81% KCl), but both 2m5-HT and 1-PBG acted as lower potency partial agonists. Ondansetron (30, 100, 300 nM) caused concentration-related rightward displacements to the concentration-effect curve to 5-HT. Nonlinear regression analysis of the effect of the ondansetron concentrations on the pEC(50) values produced a pK(B) value of 8.29 +/- 0.22. Desensitization of sensory nerves to the contractile effect of capsaicin (10 microM for 60 min) did not alter the ability of 5-HT to potentiate neurogenic contractions. 5-HT (3 microM) inhibited contractions induced by direct muscle stimulation (lignocaine, 300 microM and 10-ms pulse width). m-CPB also caused the same effect with a pIC(50) of 6.62 +/- 0.10 and an E (max) of 48.03 +/- 2.25%. The concentration-response curve to m-CPB was shifted rightwards by ondansetron (1 microM) giving an apparent pK(B) value of 8.15 +/- 0.33. mRNA for both the 5-HT(3A) and 5-HT(3B) receptor subunits was detected in the detrusor as well as the mucosa with a greater relative expression of the 5-HT(3A) subunit in both layers. This study demonstrates that 5-HT mediates enhanced neurogenic contractions of the mouse bladder muscle by an action at 5-HT(3) receptors located prejunctionally on nonsensory nerve elements. Additionally, an inhibitory postjunctional population of the 5-HT(3) receptor was identified. The presence of the 5-HT(3) receptor was confirmed by the expression of both 5-HT(3A) and 5-HT(3B) receptor subunits of the 5-HT(3) receptor.
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Department of Pharmaceutical Biology, Victorian College of Pharmacy, Monash University, Parkville, Victoria 3052, Australia.
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Chess-Williams
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Kerr
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D000818:Animals; D005260:Female; D066298:In Vitro Techniques; D051379:Mice; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects / Q000502:physiology; D017294:Ondansetron / Q000494:pharmacology; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D011985:Receptors, Serotonin / Q000096:biosynthesis / Q000235:genetics; D044406:Receptors, Serotonin, 5-HT3 / Q000096:biosynthesis / Q000502:physiology*; D012701:Serotonin / Q000494:pharmacology; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D001743:Urinary Bladder / Q000187:drug effects* / Q000502:physiology
C511230:Htr3a protein, mouse; D021122:Protein Subunits; D012333:RNA, Messenger; D011985:Receptors, Serotonin; D044406:Receptors, Serotonin, 5-HT3; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D012701:Serotonin; D017294:Ondansetron