Pei Yee Wong, Zhihui Fong, Mark A Hollywood, Keith D Thornbury, Gerard P Sergeant
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Delivery of brief (1s duration) pulses (4 Hz) yielded transient contractions that were inhibited tetrodotoxin (100 nM) and guanethidine (10 µM). α,β-meATP (10 µM), a P2X1R desensitising agent, reduced the amplitude of these responses by 65% and prazosin (100 nM), an α<sub>1</sub>-adrenoceptor antagonist, decreased mean contraction amplitude by 69%. Stimulation of α<sub>1</sub>-adrenoceptors with phenylephrine (3 µM) enhanced EFS and ATP-induced contractions and these effects were mimicked by the phorbol ester PDBu (1 µM), which activates PKC. The PKC inhibitor GF109203X (1 µM) prevented the stimulatory effects of PDBu on ATP-induced contractions of the vas deferens but only reduced the stimulatory effects of phenylephrine by 40%. PDBu increased the amplitude of ATP-induced currents recorded from freshly isolated vas deferens myocytes and HEK-293 cells expressing human P2X1Rs by 93%. 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引用次数: 0
摘要
刺激输精管中的交感神经会产生双相收缩,包括由 ATP 激活 P2X1 受体产生的快速瞬时收缩和由去甲肾上腺素激活α1-肾上腺素受体介导的次级持续收缩。去甲肾上腺素还能增强 ATP 依赖性输精管收缩,但这种作用的机制尚不清楚。本研究旨在探讨激活α1-肾上腺素受体诱导输精管瞬时收缩的潜在机制。小鼠输精管收缩是由电场刺激(EFS)诱发的。P2X1R脱敏剂α,β-meATP(10 µM)可将这些反应的幅度降低65%,α1-肾上腺素受体拮抗剂哌唑嗪(100 nM)可将平均收缩幅度降低69%。用苯肾上腺素(3 µM)刺激α1-肾上腺素受体可增强EFS和ATP诱导的收缩,而这些效应可被激活PKC的辛醇酯PDBu(1 µM)模拟。PKC 抑制剂 GF109203X(1 µM)可阻止 PDBu 对 ATP 诱导的输精管收缩的刺激作用,但只能将苯肾上腺素的刺激作用降低 40%。PDBu 使新鲜分离的输精管肌细胞和表达人 P2X1Rs 的 HEK-293 细胞记录到的 ATP 诱导电流的振幅增加了 93%。这项研究表明(1) PKC 抑制剂 GF109203X 无法完全阻断α1-肾上腺素受体激活对 ATP 诱导的小鼠输精管收缩的增效作用;(2) PKC 对 ATP 诱导的输精管收缩的刺激作用与输精管肌细胞中 P2X1R 电流的增强有关。
Regulation of nerve-evoked contractions of the murine vas deferens.
Stimulation of sympathetic nerves in the vas deferens yields biphasic contractions consisting of a rapid transient component resulting from activation of P2X1 receptors by ATP and a secondary sustained component mediated by activation of α1-adrenoceptors by noradrenaline. Noradrenaline can also potentiate the ATP-dependent contractions of the vas deferens, but the mechanisms underlying this effect are unclear. The purpose of the present study was to investigate the mechanisms underlying potentiation of transient contractions of the vas deferens induced by activation of α1-adrenoceptors. Contractions of the mouse vas deferens were induced by electric field stimulation (EFS). Delivery of brief (1s duration) pulses (4 Hz) yielded transient contractions that were inhibited tetrodotoxin (100 nM) and guanethidine (10 µM). α,β-meATP (10 µM), a P2X1R desensitising agent, reduced the amplitude of these responses by 65% and prazosin (100 nM), an α1-adrenoceptor antagonist, decreased mean contraction amplitude by 69%. Stimulation of α1-adrenoceptors with phenylephrine (3 µM) enhanced EFS and ATP-induced contractions and these effects were mimicked by the phorbol ester PDBu (1 µM), which activates PKC. The PKC inhibitor GF109203X (1 µM) prevented the stimulatory effects of PDBu on ATP-induced contractions of the vas deferens but only reduced the stimulatory effects of phenylephrine by 40%. PDBu increased the amplitude of ATP-induced currents recorded from freshly isolated vas deferens myocytes and HEK-293 cells expressing human P2X1Rs by 93%. This study indicates that: (1) potentiation of ATP-evoked contractions of the mouse vas deferens by α1-adrenoceptor activation were not fully blocked by the PKC inhibitor GF109203X and (2) that the stimulatory effect of PKC on ATP-induced contractions of the vas deferens is associated with enhanced P2X1R currents in vas deferens myocytes.
期刊介绍:
Nucleotides and nucleosides are primitive biological molecules that were utilized early in evolution both as intracellular energy sources and as extracellular signalling molecules. ATP was first identified as a neurotransmitter and later as a co-transmitter with all the established neurotransmitters in both peripheral and central nervous systems. Four subtypes of P1 (adenosine) receptors, 7 subtypes of P2X ion channel receptors and 8 subtypes of P2Y G protein-coupled receptors have currently been identified. Since P2 receptors were first cloned in the early 1990’s, there is clear evidence for the widespread distribution of both P1 and P2 receptor subtypes in neuronal and non-neuronal cells, including glial, immune, bone, muscle, endothelial, epithelial and endocrine cells.