endothelin-1 and alpha-beta-methyleneadenosine-5--triphosphate

Endothelin-1 (ET-1), an endogenous vasoconstrictor, has been known as a pro-nociceptive agent involved in multitude of pain. ET-1 acts on endothelin receptors on vascular endothelial cells, sensitizes release of ATP, which then acts on P2X3 receptors on nociceptors and results in mechanical hyperalgesia. Both endothelin receptors and P2X3 receptors are present in primary sensory neuron, where it remains unclear whether there is an interaction between them. Herein, we reported that ET-1 potentiated the electrophysiological activity of P2X3 receptors in rat dorsal root ganglia (DRG) neurons. ET-1 concentration-dependently increased α,β-methylene-ATP (α,β-meATP)-evoked inward currents, which were mediated by P2X3 receptors. ET-1 shifted the α,β-meATP concentration-response curve upwards, with an increase of 34.38 ± 4.72% in the maximal current response to α,β-meATP in the presence of ET-1. ET-1 potentiation of α,β-meATP-evoked currents was voltage-independent. ET-1 potentiated P2X3 receptor-mediated currents through endothelin-A receptors (ET

Topics: Adenosine Triphosphate; Animals; Behavior, Animal; Electrophysiological Phenomena; Endothelin-1; Ganglia, Spinal; GTP-Binding Proteins; Hyperalgesia; Male; Neurons; Protein Kinase C; Purinergic P2X Receptor Agonists; Rats; Rats, Sprague-Dawley; Receptor, Endothelin A; Receptors, Purinergic P2X3; Sensory Receptor Cells; Signal Transduction

Noradrenaline and ATP are sympathetic co-transmitters. In rat isolated mesenteric small arteries, activation of sympathetic nerves can produce a vasoconstrictor response mediated by ATP. In contrast, the rat perfused mesenteric bed displays vasoconstrictor responses that are blocked solely by alpha1-adrenoceptor antagonists. This study assessed the effect of raising tone with a vasoconstrictor on purinergic and noradrenergic responses to sympathetic nerve stimulation in the rat perfused mesentery. Rat mesenteric vascular beds were perfused with physiological salt solution and responses to nerve stimulation, or P2X-receptor agonists, were determined under basal conditions and after raising tone with endothelin-1. The contribution of noradrenaline and ATP to sympathetic nerve-mediated responses was assessed using the alpha1-adrenoceptor antagonist, prazosin and the P2X-receptor desensitizing agent, alpha,beta-methyleneATP. The effect of endothelin-1 on excitatory junction potentials generated in response to nerve stimulation in isolated mesenteric arteries was also assessed. Under baseline conditions, responses to nerve stimulation were mediated solely by activation of alpha1-adrenoceptors. After raising perfusion pressure with endothelin-1 or the thromboxane mimetic 9,11-dideoxy-11alpha,9alpha-epoxymethanoprostaglandin F2alpha (U44619), sympathetic nerve-mediated responses were larger than under basal conditions and the response was partly sensitive to P2X-receptor desensitization. Responses to exogenous P2X-receptor agonists were enhanced after treatment with endothelin-1, while endothelin-1 decreased the amplitude of excitatory junction potentials. These results indicate that ATP acts as an important, functional, sympathetic neurotransmitter in the perfused mesentery under raised tone conditions, where the perfusion pressure is closer to that found in vivo. This effect is due to a postjunctional enhancement of purinergic function.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Adenosine Triphosphate; Adrenergic alpha-Antagonists; Animals; Dose-Response Relationship, Drug; Electric Stimulation; Endothelin-1; In Vitro Techniques; Male; Mesenteric Arteries; Norepinephrine; Prazosin; Rats; Rats, Wistar; Receptors, Adrenergic, alpha-1; Receptors, Purinergic P2; Receptors, Purinergic P2X; Splanchnic Circulation; Sympathetic Nervous System; Vascular Resistance; Vasoconstriction; Vasoconstrictor Agents

We aimed to compare the expression and function of molecular components of the RhoA/Rho-kinase signaling pathway in the contractile responses of detrusor, trigonal and urethral smooth muscle, using selective Rho-kinase inhibitors. Contractility studies and molecular approaches were employed to demonstrate the expression patterns and functional activity of the RhoA/Rho-kinase signaling pathway in the lower urinary tract. Frequency-response curves (1-32 Hz) and concentration-response curves (CRC) to carbachol (CCh, 0.01-30 microM), phenylephrine (PE, 0.01-300 microM) and endothelin-1 (ET-1, 0.01-100 nM) were significantly attenuated (p<0.01) following incubation with the Rho-kinase inhibitors H-1152 (0.1-1 microM), Y-27632 (1-10 microM) or HA-1077 (10 microM). Addition of Rho-kinase inhibitors also markedly reduced (p<0.01) the contractions evoked by either KCl (80 mM) or alpha,beta-methylene ATP (alpha,beta-mATP, 10 microM). Among the Rho-kinase inhibitors tested, H-1152 was approximately 9-16 times more potent than Y-27632 or HA-1077. In addition, basal tone of detrusor and trigonal strips was reduced following addition of Y-27632 (10 microM), H-1152 (1 microM) and HA-1077 (10 microM). The expression of RhoA, RhoGDI, leukemia-associated RhoGEF (LARG) and p115RhoGEF was similar among the detrusor, trigone and urethra, whereas Rho-kinase alpha, Rho-kinase beta and PDZ-RhoGEF protein levels were significantly lower in the urethra. Components of the RhoA/Rho-kinase signaling are expressed in detrusor, trigonal and urethral smooth muscle and dynamically regulate contraction and tone. Manipulation of RhoGEF expression may provide further understanding of mechanisms involving Ca(2+) sensitization in the lower urinary tract.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Adenosine Triphosphate; Amides; Animals; Atropine; Blotting, Western; Calcium Signaling; Dose-Response Relationship, Drug; Electric Stimulation; Endothelin-1; Enzyme Inhibitors; Guanine Nucleotide Dissociation Inhibitors; In Vitro Techniques; Intracellular Signaling Peptides and Proteins; Male; Muscle Contraction; Muscle, Smooth; Nifedipine; Protein Serine-Threonine Kinases; Pyridines; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; rho-Associated Kinases; rho-Specific Guanine Nucleotide Dissociation Inhibitors; rhoA GTP-Binding Protein; Urethra; Urinary Bladder