Location-dependent sensing of nitric oxide and calcium ions in living rat kidney using an amperometric/potentiometric dual microsensor.
Location-dependent sensing of nitric oxide and calcium ions in living rat kidney using an amperometric/potentiometric dual microsensor.
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DOI：
10.1039/c5an01804h
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发表时间：
2016-01
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影响因子：
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通讯作者：
Yee‐Seul Kim;Yejin Ha;Jung-Bo Sim;M. Suh;Youngmi Lee
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文献类型：
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作者：
Yee‐Seul Kim;Yejin Ha;Jung-Bo Sim;M. Suh;Youngmi Lee
本文报道了一种用于检测一氧化氮（NO）和钙离子（Ca(2+)）的双微传感器的制备及其在活体大鼠肾组织中同时测量NO/Ca（2+）的应用。NO和Ca（2+）具有非常重要的生理功能，并且都复杂地参与了许多生物过程。基于Pt（直径25 μm）和Ag（直径76 μm）微盘的双凹槽电极制备了双NO/Ca（2+）传感器。在铂盘面（WE1）表面电沉积多孔铂黑，然后涂上氟化干凝胶；用于NO的安培传感。将银盘表面（WE2）氯化成AgCl，然后进行硅烷化，最后进行Ca（2+）选择性膜负载；用于Ca（2+）电位感应。该双传感器对NO具有高灵敏度（40.8±6.5 pA μM(-1), n = 10），对Ca（2+）变化具有可靠的Nerntian响应（25.7±0.5 mV pCa(-1), n = 10），对NO和Ca（2+）的选择性优于常见干扰，稳定性可靠（可达4 h组织实验）。所制备的传感器用于实时监测10 mM l-N(G)-硝基精氨酸甲酯（l-NAME， NO合酶抑制剂）诱导大鼠肾脏NO和Ca（2+）水平的动态变化。由于传感器尺寸小，NO和Ca（2+）的位置依赖分析是在肾脏的两个不同区域（肾髓质和皮质）进行的。髓质的NO和Ca（2+）水平高于皮质。本研究验证了实时监测密切相关的Ca（2+）和内源NO生成的可行性；以及局部NO和Ca（2+）的浓度评估。
In this paper, we report the fabrication of a dual microsensor for sensing nitric oxide (NO) and calcium ions (Ca(2+)) and its application for simultaneous NO/Ca(2+) measurements in living rat kidney tissue. NO and Ca(2+) have very important physiological functions and are both intricately involved in many biological processes. The dual NO/Ca(2+) sensor is prepared based on a dual recessed electrode possessing Pt (diameter, 25 μm) and Ag (diameter, 76 μm) microdisks. The Pt disk surface (WE1) is electrodeposited with porous Pt black and then coated with fluorinated xerogel; and used for amperometric sensing of NO. The Ag disk surface (WE2) is chloridated to AgCl, followed by silanization and then Ca(2+) selective membrane loading; and used for potentiometric sensing of Ca(2+). The dual sensor exhibits high sensitivity of WE1 to NO (40.8 ± 6.5 pA μM(-1), n = 10) and reliable Nerntian response of WE2 to Ca(2+) changes (25.7 ± 0.5 mV pCa(-1), n = 10) with excellent selectivity to only NO and Ca(2+) over common interferents and reliable stability (up to ∼4 h tissue experiment). The prepared sensor is employed for real-time monitoring of the dynamic changes of NO and Ca(2+) levels of a rat kidney, which is induced by the administration of 10 mM l-N(G)-nitroarginine methyl ester (l-NAME, a NO synthase inhibitor). Due to the small sensor dimension, location-dependent analyses of NO and Ca(2+) are carried out at two different regions of a kidney (renal medulla and cortex). Higher NO and Ca(2+) levels are observed at the medulla than at the cortex. This study verifies the feasibility for real-time monitoring of intimately connected Ca(2+) and endogenous NO production; and also for localized concentration assessments of both NO and Ca(2+).