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首页> 外文期刊>American Journal of Physiology >P2X purinergic receptor channel expression and function in bovine aortic endothelium.
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P2X purinergic receptor channel expression and function in bovine aortic endothelium.

机译：P2X嘌呤能受体通道在牛主动脉内皮中的表达和功能。

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摘要

We examined bovine aortic endothelial cells (BAECs) for the functional expression of P2X receptors, the ATP-gated cation channels. We identified the P2X subtypes present in BAECs using RT-PCR. mRNA was present for only three of seven family members: P2X4, P2X5, and P2X7. We then characterized agonist-activated currents in whole cell and outside-out patch recordings using 2-methyl-thio-ATP (MeSATP) as a P2X4 and P2X5 receptor agonist and 2',3'-O-(4-benzoylbenzoyl)ATP (BzATP) as a P2X7 receptor agonist. MeSATP (10-20 microM) produced current with characteristics of P2X4 receptors. The current was an inwardly rectifying current, reversed near 0 mV, slowly desensitized, was not blocked by suramin (300 microM) or reactive blue (60 microM), and had a single channel conductance of 36 pS. BzATP (10-100 microM), on the other hand, activated a 9-pS channel with sustained activity in the continued presence of the agonist. BzATP-activated current was blocked by reactive blue (60 microM) and by suramin (approximately 50% block at 300 microM). We confirmed, by immunocytochemistry, the presence of P2X4 and P2X7 protein. The agonists failed, however, to induce significant uptake of the large molecule YO-PRO, indicating the lack of pore development that has been demonstrated for P2X7 and P2X4 in response to agonist in some cell types.
机译：我们检查了牛主动脉内皮细胞（BAEC）的P2X受体，ATP门控阳离子通道的功能性表达。我们使用RT-PCR鉴定了BAEC中存在的P2X亚型。只有七个家族成员中的三个存在mRNA：P2X4，P2X5和P2X7。然后，我们使用2-甲基-硫代ATP（MeSATP）作为P2X4和P2X5受体激动剂和2'，3'-O-（4-苯甲酰基苯甲酰基）ATP（ BzATP）作为P2X7受体激动剂。 MeSATP（10-20 microM）产生具有P2X4受体特征的电流。该电流是向内的整流电流，在0 mV左右反向，缓慢脱敏，未被苏拉明（300 microM）或活性蓝（60 microM）阻断，单通道电导为36 pS。另一方面，在持续存在激动剂的情况下，BzATP（10-100 microM）激活了具有持续活性的9-pS通道。 BzATP激活的电流被活性蓝（60 microM）和苏拉明（苏拉明）（在300 microM时约50％的阻断）阻断。我们通过免疫细胞化学证实了P2X4和P2X7蛋白的存在。但是，激动剂未能诱导大分子YO-PRO的大量摄取，表明缺乏对P2X7和P2X4在某些细胞类型中对激动剂的反应所证实的孔发展。

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