Two-photon imaging of endothelin-1-mediated intracellular Ca(2+) handling in smooth muscle cells of rat renal resistance arteries.
Two-photon imaging of endothelin-1-mediated intracellular Ca(2+) handling in smooth muscle cells of rat renal resistance arteries.
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DOI：
10.1016/j.lfs.2015.12.022
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发表时间：
2016-08-15
期刊：
影响因子：
6.1
通讯作者：
Staruschenko A
中科院分区：
文献类型：
--
作者：
Palygin O;Miller B;Ilatovskaya DV;Sorokin A;Staruschenko A
内皮素-1（ET-1）是一种强有力的血管收缩剂，调节心肾系统的生理功能。本研究的目的是评估ET-1介导的肾阻力动脉平滑肌细胞（SMC）内钙升高。在原代SMC的体外研究中，从大鼠肾微血管中分离，细胞内Ca 2+的水平计算与Fura 2-AM染料加载细胞后，在340和380 nm的发射率。在离体研究中，我们使用双光子成像的细胞内Ca 2+处理肾阻力动脉切除大鼠肾脏和加载荧光Ca 2+指示剂Fluo-4 AM。双光子成像显示，用ET-1处理离体大鼠肾阻力动脉导致这些血管平滑肌血管中细胞内Ca 2+浓度迅速增加。这些离体观察结果与体外研究结果一致，表明ET-1介导TRPC通道的激活，并将培养的SMC中的细胞内Ca 2+水平增加至510±83 nM。ET-1介导的细胞内Ca 2+升高与肾微血管收缩密切相关，并且对于ET-1诱导的SMC收缩至关重要。双光子成像的细胞内钙在大鼠肾阻力动脉的完整的SMC是一个强大的技术，它允许详细的离体分析细胞内钙处理ET-1，在高血压相关的肾脏疾病的重要球员。
Endothelin-1 (ET-1) is a potent vasoconstrictor which regulates the physiology of cardiorenal system. The aim of this study was to evaluate ET-1-mediated elevation of intracellular Ca2+ in smooth muscle cells (SMC) of renal resistance arteries. In in vitro studies of primary SMC, which were isolated from rat renal microvessels, the levels of intracellular Ca2+ were calculated from the ratio of emissions at 340 and 380 nm after loading cells with Fura 2-AM dye. In ex vivo studies we used two-photon imaging of intracellular Ca2+ handling in renal resistance arteries excised from rat kidneys and loaded with fluorescent Ca2+ indicator Fluo-4 AM. The two-photon imaging demonstrates that treatment of isolated rat renal resistance arteries with ET-1 causes a rapid increase of intracellular Ca2+ concentration in smooth muscle vasculature of these vessels. These ex vivo observations are in accordance with in vitro findings indicating that ET-1 mediates activation of TRPC channels and increases the level of intracellular Ca2+ in cultured SMC to 510±83 nM. ET-1-mediated elevation of intracellular Ca2+ is strongly linked to renal microvascular contraction and is crucial for ET-1-induced contraction of SMC. The two-photon imaging of intracellular Ca2+ in intact SMC of rat renal resistance arteries is a powerful technique which allows the detailed ex vivo analysis of intracellular Ca2+ handling by ET-1, an important player in hypertension-related kidney diseases.