Purinergic signaling underlies CFTR control of human airway epithelial cell volume.
Purinergic signaling underlies CFTR control of human airway epithelial cell volume.
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DOI：
10.1016/j.jcf.2004.01.006
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发表时间：
2004-06-01
期刊：
影响因子：
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通讯作者：
Schwiebert, Erik M
中科院分区：
文献类型：
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作者：
Braunstein, Gavin M;Zsembery, Akos;Schwiebert, Erik M
背景：囊性纤维化（CF）中囊性纤维化跨膜传导调节因子（CFTR）功能的丧失导致上皮中多种离子通道、水通道和酸碱转运蛋白的失调。因此，我们假设许多关键离子通道和转运体的失调可能导致人类气道上皮细胞体积调节的缺陷。方法：通过Coulter Counter Multisizer iii驱动的电子细胞定格，实时评估非CF、CF和cftr补充的CF人气道上皮细胞的细胞体积、调节体积减少及其调节作用。SPQ卤化物荧光测定低渗诱导的氯化物外排提供了细胞体积测定的间接验证。结果：CFTR通过自分泌ATP信号调控人气道上皮细胞体积。非CF细胞和野生型(WT)- cftr转染的CF细胞有正常的调节体积减少（RVD）反应，这种反应通过阻断自分泌和旁分泌嘌呤能信号而减弱。相比之下，亲代IB3-1 CF细胞或表达CFTR突变体（DeltaF508、G551D和S1455X）的IB3-1细胞未能发生RVD。CF细胞的RVD可通过P2Y G蛋白偶联受体的激动剂和P2X嘌呤能受体通道的激动剂来挽救。结论：CFTR和CFTR驱动的自分泌ATP信号的缺失可能是CF气道上皮细胞体积调节缺陷和离子、水和酸碱运输失调的基础。
BACKGROUND: Loss of cystic fibrosis transmembrane conductance regulator (CFTR) function in cystic fibrosis (CF) causes dysregulation of multiple ion channels, water channels, and acid-base transporters in epithelia. As such, we hypothesized that dysregulation of many critical ion channels and transporters may cause defects in human airway epithelial cell volume regulation.METHODS: Cell volume, regulatory volume decrease, and its regulation was assessed in real-time via Coulter Counter Multisizer III-driven electronic cell sizing in non-CF, CF, and CFTR-complemented CF human airway epithelial cells. SPQ halide fluorescence assay of hypotonicity-induced chloride efflux provided indirect validation of the cell volume assays.RESULTS: CFTR, via autocrine ATP signaling, governs human airway epithelial cell volume regulation. Non-CF cells and wild-type (WT)-CFTR-transfected CF cells had normal regulatory volume decrease (RVD) responses that were attenuated by blockade of autocrine and paracrine purinergic signaling. In contrast, parental IB3-1 CF cells or IB3-1 cells expressing CFTR mutants (DeltaF508, G551D, and S1455X) failed to RVD. CF cell RVD was rescued by agonists to P2Y G protein-coupled receptors and, more robustly, by agonists to P2X purinergic receptor channels.CONCLUSIONS: Loss of CFTR and CFTR-driven autocrine ATP signaling may underlie defective cell volume regulation and dysregulated ion, water, and acid-base transport in CF airway epithelia.