Detection of nitric oxide release from single neurons in the pond snail, Lymnaea stagnalis
Detection of nitric oxide release from single neurons in the pond snail, Lymnaea stagnalis
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DOI：
10.1021/ac060863w
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发表时间：
2006-11-15
影响因子：
7.4
通讯作者：
O'Hare, Danny
中科院分区：
文献类型：
--
作者：
Patel, Bhavik Anil;Arundell, Martin;O'Hare, Danny
使用 Nafion 和电聚合聚丁香酚或邻苯二胺在 30 微米碳纤维圆盘电极上制造了多个薄膜涂层一氧化氮传感器。这是一项罕见的研究，它利用圆盘电极而不是广泛使用的突出尖端微电极来从生物环境中进行测量。这些电极已用于评估池塘蜗牛（Lymnaea stagnalis）中两个不同的已识别神经元之间一氧化氮释放的差异。这些结果首次直接测量了单个神经元释放的一氧化氮。该电极对一氧化氮非常敏感，检测限为 2.8 nM，灵敏度为 9.46 nA mu M-1。该传感器对多种神经化学干扰物（如抗坏血酸、尿酸和儿茶酚胺）以及二次氧化产物（如亚硝酸盐）具有很强的选择性。在完整但孤立的中枢神经系统中，测量了两个神经元、脑巨细胞 (CGC) 和 B2 颊运动神经元的细胞体释放的一氧化氮。高 Ca2+/高 K+ 刺激能够引起可重复的释放。对于给定的刺激，B2 神经元比 CGC 神经元释放更多的一氧化氮；然而，两种细胞均受到 NOS 抑制剂 L-NAME 的抑制。
Multiple film-coated nitric oxide sensors have been fabricated using Nafion and electropolymerized polyeugenol or o-phenylenediamine on 30-mu m carbon fiber disk electrodes. This is a rare study that utilizes disk electrodes rather than the widely used protruding tip microelectrodes in order to measure from a biological environment. These electrodes have been used to evaluate the differences in nitric oxide release between two different identified neurons in the pond snail, Lymnaea stagnalis. These results show the first direct measurements of nitric oxide release from individual neurons. The electrodes are very sensitive to nitric oxide with a detection limit of 2.8 nM and a sensitivity of 9.46 nA mu M-1. The sensor was very selective against a variety of neurochemical interferences such as ascorbic acid, uric acid, and catecholamines and secondary oxidation products such as nitrite. Nitric oxide release was measured from the cell bodies of two neurons, the cerebral giant cell (CGC) and the B2 buccal motor neuron, in the intact but isolated CNS. A high-Ca2+/high-K+ stimulus was capable of evoking reproducible release. For a given stimulus, the B2 neuron released more nitric oxide than the CGC neuron; however, both cells were equally suppressed by the NOS inhibitor L-NAME.