Shinohara Hiroaki, Wang Feifei
Field of Life, Information and System Sciences, Graduate School of Science and Engineering, University of Toyama, Japan.
Anal Sci. 2007 Jan;23(1):81-4. doi: 10.2116/analsci.23.81.
A real-time observation of neurotransmitter release from a nerve cell is a useful method for not only neuroscience research, but also assessing of the influence of chemicals, including drugs, on the human nervous system. In this study, a more simple and sensitive method for real-time monitoring of dopamine release from a nerve model cell was developed. Highly sensitive detection of dopamine was performed by using tyramine oxidase for dopamine oxidation, which was followed by a luminol luminescence reaction. This enzyme-catalyzed luminescence method was applied to observe dopamine release from the PC12 cell as a nerve model cell upon stimulation with acetylcholine and an acetylcholine receptor agonist. The results demonstrated that the real-time monitoring of the activation of the PC12 cell was easily performed by this method. This method possessed many advantages, such as high sensitivity, rapid measurement and no pretreatment for cells. It might be applied to drug screening and the assessment of harmful influences of food additives and pesticides on the nerves.
对神经细胞神经递质释放进行实时观察，不仅是神经科学研究的一种有用方法，也是评估包括药物在内的化学物质对人类神经系统影响的一种有用方法。在本研究中，开发了一种更简单、灵敏的实时监测神经模型细胞多巴胺释放的方法。通过使用酪胺氧化酶氧化多巴胺，随后进行鲁米诺发光反应，实现了对多巴胺的高灵敏度检测。将这种酶催化发光方法应用于观察作为神经模型细胞的PC12细胞在乙酰胆碱和乙酰胆碱受体激动剂刺激下的多巴胺释放。结果表明，通过该方法可以轻松实现对PC12细胞激活的实时监测。该方法具有许多优点，如灵敏度高、测量快速且无需对细胞进行预处理。它可能适用于药物筛选以及评估食品添加剂和农药对神经的有害影响。
