Development of capillary-based electrochemical sensors for bioactive substances in brain

开发基于毛细管的脑生物活性物质电化学传感器

• 批准号: 12440210
• 负责人: SUGAWARA Masao
• 金额: $ 8.38万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12440210/
• 关键词: biomembrane sensors; electrochemical sensors; bilayer lipid membrane; neurotransmitter; L-glutamate; arachidonic acid; mouse hippocampus; enzyme sensor; ガラスキャピラリー; 超微小電極; マウス脳海馬; グラミシジン; 平面脂質二分子膜; 電気分析法; 動電的イオンサンプリング能

1. Two kinds of glass capillary sensors, one with the tip diameter of several μm for electrokinetic sampling of analyte ions and one with the tip diameter of 〜10 μm for sampling of analytes based on capillarity, have been developed applied to detection of L-glutamate released in mouse brain slices.2. It was found that arachidonic acid induces a channel-type current through selectively interaction with a planar bilayer lipid membrane. The arachidonic acid-induced channel current was useful as analytical signal that reflects arachidonic acid concentration.3. Bilayer lipid membrane sensors based on a membrane bound separate receptors such as biotin-PE and DNP-PE at the membrane interface induced modulation of the open/close kinetics of a gramicidin channel, which can be used as a measure of analyte concentration.4. Microsensors based on L-glutamate receptor subtypes embedded in bilayer lipid membranes were constructed for evaluation of agonist selectivity based on ion permeation ability. The agonist selectivity based on ion permeation was found not parallel to the binding affinity of the agonists to the receptor.

1.研制了两种玻璃毛细管传感器，一种用于电动采样，针尖直径为几μm，另一种用于毛细管采样，针尖直径为10 μm，并将其应用于小鼠脑片中L-谷氨酸的检测.结果发现，花生四烯酸通过选择性地与平面双层脂质膜相互作用诱导通道型电流。花生四烯酸诱导的通道电流可作为反映花生四烯酸浓度的分析信号.基于膜结合的双层脂质膜传感器在膜界面处诱导短杆菌肽通道的开/闭动力学的调节，其可以用作分析物浓度的测量，所述膜结合的分离受体例如生物素-PE和DNP-PE。基于L-谷氨酸受体亚型嵌入双层脂膜的微传感器的构建，用于基于离子渗透能力的激动剂选择性评价。发现基于离子渗透的激动剂选择性与激动剂对受体的结合亲和力不平行。

项目成果

A.Hirano: "Evaluation of agonist selectivity for the NMDA receptor ion channel in bilayer lipid membranes based on integrated single-channel currents"Biosensors & Bioelectronic. 15. 173-181 (2000)

A.Hirano：“基于集成单通道电流评估双层脂膜中 NMDA 受体离子通道的激动剂选择性”生物传感器

M.Sugawara: "Design and Application of Ion-Channel Sensors Based on Biological and Artificial Receptors"Bull.Chem.Soc.Jp. 75. 187-201 (2002)

M.Sukawara：“基于生物和人工受体的离子通道传感器的设计与应用”Bull.Chem.Soc.Jp。

菅原 正雄: "キャピラリー電気化学センサーの開発と応用"分析化学. 51. 1121-1133 (2002)

Masao Sukawara：“毛细管电化学传感器的开发和应用”分析化学51。1121-1133（2002）。

A.Hirano: "A single-channel sensor based on gramicidin controlled by molecular recognition at bilayer lipid membranes containing receptor"Biosensors and Bioelectronics. (In press). (2003)

A.Hirano：“一种基于短杆菌肽的单通道传感器，由含有受体的双层脂膜上的分子识别控制”生物传感器和生物电子学。

N.Shimokawa: "An Ion-Channel Sensot for Abasic Sites in DNA"Anal.Sci.. 17. i1379-i1382 (2001)

N.Shimokawa：“DNA 中无碱基位点的离子通道传感器”Anal.Sci.. 17. i1379-i1382 (2001)