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首页> 美国卫生研究院文献>British Journal of Pharmacology and Chemotherapy >Lack of run-down of smooth muscle P2X receptor currents recorded with the amphotericin permeabilized patch technique physiological and pharmacological characterization of the properties of mesenteric artery P2X receptor ion channels
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Lack of run-down of smooth muscle P2X receptor currents recorded with the amphotericin permeabilized patch technique physiological and pharmacological characterization of the properties of mesenteric artery P2X receptor ion channels

机译：用两性霉素透化贴片技术记录的平滑肌P2X受体电流缺乏肠系膜动脉P2X受体离子通道的生理和药理学表征

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class="enumerated" style="list-style-type:decimal">Immunoreactivity for P2X1, P2X4 and P2X5 receptor subtypes was detected in the smooth muscle cell layer of second and third order rat mesenteric arteries immunoreactivity, for P2X2, P2X3, P2X6 and P2X7 receptors was below the level of detection in the smooth muscle layer.P2X receptor-mediated currents were recorded in patch clamp studies on acutely dissociated mesenteric artery smooth muscle cells. Purinergic agonists evoked transient inward currents that decayed rapidly in the continued presence of agonist (τ∼200 ms). Standard whole cell responses to repeated applications of agonist at 5 min intervals ran down. Run-down was unaffected by changes in extracellular calcium concentration, intracellular calcium buffering or the inclusion of ATP and GTP in the pipette solution.Run-down was overcome and reproducible responses to purinergic agonists were recorded using the amphotericin permeabilized patch recording configuration.The rank order of potency at the P2X receptor was ATP=2 methylthio ATP>α,β-methylene ATP>CTP=l-β,γ-methylene ATP. Only ATP and 2meSATP were full agonists. The P2 receptor antagonists suramin and PPADS inhibited P2X receptor-mediated currents with IC50s of 4 μM and 70 nM respectively.These results provide further characterization of artery P2X receptors and demonstrate that the properties are dominated by a P2X1-like receptor phenotype. No evidence could be found for a phenotype corresponding to homomeric P2X4 or P2X5 receptors or to heteromeric P2X1/5 receptors and the functional role of these receptors in arteries remains unclear.
机译：class =“ enumerated” style =“ list-style-type：decimal”> <！-list-behavior =枚举前缀-word = mark-type = decimal max-label-size = 0-> 在二，三阶大鼠肠系膜动脉的免疫细胞中，对P2X1，P2X4和P2X5受体亚型具有免疫反应性，其中P2X2，P2X3，P2X6和P2X7受体低于在平滑肌层中的检测水平。在膜片钳研究中记录了急性解离的肠系膜动脉平滑肌细胞的li> liP2X受体介导的电流。嘌呤能激动剂引起瞬态内向电流，在持续存在激动剂（τ〜200μms）时，其迅速衰减。标准全细胞对以5分钟间隔重复使用激动剂的反应下降了。移液不受细胞外钙浓度变化，细胞内钙缓冲或移液器溶液中ATP和GTP含量的影响。 克服了移液现象，并使用两性霉素记录了对嘌呤能激动剂的可再现反应 P2X受体的效能等级顺序为ATP = 2甲硫基ATP>α，β-亚甲基ATP> CTP =1-β，γ-亚甲基ATP。只有ATP和2meSATP是完全激动剂。 P2受体拮抗剂苏拉明和PPADS抑制P2X受体介导的电流，IC50分别为4μM和70 nM。 这些结果提供了对动脉P2X受体的进一步表征，并证明其特性主要由P2X1决定。像受体表型。没有证据表明对应于同型P2X4或P2X5受体或异型P2X1 / 5受体的表型，这些受体在动脉中的功能作用仍不清楚。

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