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首页> 外文期刊>American Journal of Physiology >Transepithelial pressure pulses induce nucleotide release in polarized MDCK cells.
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Transepithelial pressure pulses induce nucleotide release in polarized MDCK cells.

机译：经上皮压力脉冲诱导极化的MDCK细胞中的核苷酸释放。

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摘要

The release of nucleotides is involved in mechanosensation in various epithelial cells. Intriguingly, kidney epithelial cells are absolutely dependent on the primary cilium to sense changes in apical laminar flow. During fluid passage, the renal epithelial cells are subjected to various mechanical stimuli in addition to changes in the laminar flow rate. In the distal part of the collecting duct, the epithelial cells are exposed to pressure changes and possibly distension during papillary contractions. The aim of the present study was to determine whether nucleotide release contributes to mechanosensation in kidney epithelial cells, thereby establishing whether pressure changes are sufficient to produce nucleotide-mediated responses. Madin-Darby canine kidney (MDCK) cells grown on permeable supports were mounted in a closed double perfusion chamber on an inverted microscope. The intracellular Ca(2+) concentration ([Ca(2+)](i)) was monitored with the Ca(2+)-sensitive fluorescence probe fluo 4. Transepithelial pressure pulses of 30-80 mm Hg produced a transient increase in [Ca(2+)](i) of MDCK cells. This response is independent of the primary cilium, since it is readily observed in immature cells that do not yet express primary cilia. The amplitudes of the pressure-induced Ca(2+) transients varied with the applied chamber pressure in a quantity-dependent manner. The ATPase apyrase and the P2Y antagonist suramin significantly reduced the pressure-induced Ca(2+) transients. Applying apyrase or suramin to both sides of the preparation simultaneously nearly abolished the pressure-induced Ca(2+) response. In conclusion, these observations suggest that rapid pressure changes induce both apical and basolateral nucleotide release that contribute to mechanosensation in kidney epithelial cells.
机译：核苷酸的释放与各种上皮细胞的机械感觉有关。有趣的是，肾上皮细胞绝对依赖于初级纤毛来感知顶端层流的变化。在流体通过期间，除了层流速率的变化之外，肾上皮细胞还受到各种机械刺激。在收集导管的远端，上皮细胞在乳头状收缩过程中会受到压力变化的影响，并可能会膨胀。本研究的目的是确定核苷酸释放是否有助于肾脏上皮细胞的机械感觉，从而确定压力变化是否足以产生核苷酸介导的反应。将在可渗透支持物上生长的Madin-Darby犬肾（MDCK）细胞安装在倒置显微镜的密闭双灌注室中。用Ca（2+）敏感的荧光探针fluo 4监测细胞内Ca（2+）的浓度（[Ca（2 +）]（i））。30-80 mm Hg的跨上皮压力脉冲产生瞬时增加。 MDCK细胞的[Ca（2 +）]（i）。该反应与初级纤毛无关，因为它在尚未表达初级纤毛的未成熟细胞中很容易观察到。压力诱导的Ca（2+）瞬变的幅度随所施加的腔室压力以数量依赖的方式变化。 ATPase腺苷三磷酸酶和P2Y拮抗剂苏拉明显着降低了压力诱导的Ca（2+）瞬变。同时将腺苷三磷酸双磷酸酶或苏拉明应用于准备的两面几乎消除了压力诱导的Ca（2+）反应。总之，这些观察结果表明，快速的压力变化会诱导顶突和基底外侧核苷酸释放，这有助于肾脏上皮细胞的机械感受。

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