Activation of P2Y receptors causes strong and persistent shrinkage of C11-MDCK renal epithelial cells.
Activation of P2Y receptors causes strong and persistent shrinkage of C11-MDCK renal epithelial cells.
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P2Y 受体的激活导致 C11-MDCK 肾上皮细胞强烈且持续的收缩。
DOI：
10.1152/ajpcell.00018.2011
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Orlov,SergeiN
中科院分区：
文献类型：
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作者：
Koltsova,SvetlanaV;Platonova,Alexandra;Maksimov,GeorgyV;Mongin,AlexanderA;Grygorczyk,Ryszard;Orlov,SergeiN
嘌呤能受体激活多种信号级联并调节细胞体积敏感性离子转运蛋白的活性。然而，ATP和其他P2受体激动剂对细胞体积动力学的影响几乎没有研究。在目前的工作中，我们使用了最近开发的双图像表面重建技术，探讨嘌呤能激动剂对细胞体积的影响，在C11-Madin-Darby犬肾细胞系类似于肾集合管的闰细胞。出乎意料的是，我们发现ATP和UTP引发了非常强烈的（55-60%）细胞收缩，在激动剂洗脱后持续长达2小时。细胞体积的嘌呤能调节需要细胞内Ca 2+的增加，并且可以通过Ca 2+离子载体离子霉素或通过4β-佛波醇12-肉豆蔻酸酯13-乙酸酯激活蛋白激酶C来部分模拟。细胞收缩伴随着细胞内K+和Cl−含量的强烈降低，使用稳态86 Rb+和36 Cl −分布测量。收缩和离子流出在ATP处理的细胞被阻止的阴离子通道阻断剂5-硝基-2-（3-苯基丙基氨基）苯甲酸（NPPB）和BKC通道抑制剂charybdotoxin，iberotoxin，和paxilline。为了评估细胞体积变化在嘌呤能信号传导中的意义，我们测量了ATP对即刻早期基因c-Fos表达的影响。用ATP处理30分钟使c-Fos免疫反应性增加约5倍，这种作用被Charybdotoxin强烈抑制，并被NPPB完全阻止。总体而言，我们的研究结果表明，ATP诱导的细胞体积的变化是部分负责嘌呤激动剂的生理作用。
Purinergic receptors activate diverse signaling cascades and regulate the activity of cell volume-sensitive ion transporters. However, the effects of ATP and other agonists of P2 receptors on cell volume dynamics are only scarcely studied. In the present work, we used the recently developed dual-image surface reconstruction technique to explore the influence of purinergic agonists on cell volume in the C11-Madin-Darby canine kidney cell line resembling intercalated cells from kidney collecting ducts. Unexpectedly, we found that ATP and UTP triggered very robust (55–60%) cell shrinkage that lasted up to 2 h after agonist washout. Purinergic regulation of cell volume required increases in intracellular Ca2+and could be partially mimicked by the Ca2+-ionophore ionomycin or activation of protein kinase C by 4β-phorbol 12-myristate 13-acetate. Cell shrinkage was accompanied by strong reductions in intracellular K+and Cl−content measured using steady-state86Rb+and36Cl−distribution. Both shrinkage and ion efflux in ATP-treated cells were prevented by the anion channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) and by the BKCachannel inhibitors charybdotoxin, iberiotoxin, and paxilline. To evaluate the significance of cell-volume changes in purinergic signaling, we measured the impact of ATP on the expression of the immediate-early gene c-Fos. Thirty-minute treatment with ATP increased c-Fos immunoreactivity by approximately fivefold, an effect that was strongly inhibited by charybdotoxin and completely prevented by NPPB. Overall, our findings suggest that ATP-induced cell-volume changes are partially responsible for the physiological actions of purinergic agonists.