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大鼠颈动脉体化学感受器细胞缺氧诱发ATP释放的检测

Detection of hypoxia-evoked ATP release from chemoreceptor cells of the rat carotid body.

作者信息

Buttigieg Josef, Nurse Colin A

机构信息

Department of Biology, McMaster University, Hamilton, Ont., Canada L8S 4K1.

出版信息

Biochem Biophys Res Commun. 2004 Sep 10;322(1):82-7. doi: 10.1016/j.bbrc.2004.07.081.

Abstract

The carotid body (CB) is a chemosensory organ that detects changes in chemical composition of arterial blood and maintains homeostasis via reflex control of ventilation. Thus, in response to a fall in arterial PO(2) (hypoxia), CB chemoreceptors (type I cells) depolarize, and release neurotransmitters onto afferent sensory nerve endings. Recent studies implicate ATP as a key excitatory neurotransmitter released during CB chemoexcitation, but direct evidence is lacking. Here we use the luciferin-luciferase bioluminescence assay to detect ATP, released from rat chemoreceptors in CB cultures, fresh tissue slices, and whole CB. Hypoxia evoked an increase in extracellular ATP, that was inhibited by L-type Ca(2+)channel blockers and reduced by the nucleoside hydrolase, apyrase. Additionally, iberiotoxin (IbTX; 100 nM), a blocker of O(2)-sensitive Ca(2+)-dependent K(+) (BK) channels, stimulated ATP release and largely occluded the effect of hypoxia. These data strongly support a neurotransmitter role for ATP in carotid body function.

摘要

颈动脉体(CB)是一种化学感受器官,可检测动脉血化学成分的变化,并通过通气反射控制维持体内平衡。因此,响应于动脉血氧分压(PO₂)下降(缺氧),CB化学感受器(I型细胞)去极化,并将神经递质释放到传入感觉神经末梢上。最近的研究表明,ATP是CB化学兴奋过程中释放的一种关键兴奋性神经递质,但缺乏直接证据。在这里,我们使用荧光素-荧光素酶生物发光测定法来检测从CB培养物、新鲜组织切片和完整CB中的大鼠化学感受器释放的ATP。缺氧引起细胞外ATP增加,L型钙通道阻滞剂可抑制这种增加,核苷水解酶、腺苷三磷酸双磷酸酶可使其减少。此外,埃博毒素(IbTX;100 nM),一种对氧敏感的钙依赖性钾(BK)通道阻滞剂,刺激ATP释放,并在很大程度上消除了缺氧的影响。这些数据有力地支持了ATP在颈动脉体功能中作为神经递质的作用。

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