权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

ELECTROPHYSIOLOGICAL ANALYSIS OF THE SYNAPTIC ACTIVITY OF NONSPIKING INTERNEURONS IN BEHAVING ANIMALS USING WHOLE CELL RECORDING TECHNIQUES

使用全细胞记录技术对行为动物非尖峰中间神经元突触活动进行电生理分析

基本信息

批准号：
09440274
负责人：
TAKAHATA Masakazu
金额：
$ 6.66万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

In order to consolidate the experimental background for applying the whole-cell recording technique to nonspiking interneurons in the central nervous system of crayfish, Procambarus clarkii Girard, performing behavioral acts, I have focused this study on 1) new development of an unanesthetized whole animal preparation to which micro- and patch electrodes could be applicable, 2) electrophysiological analyses of membrane current of nonspiking interneurons using the single electrode voltage clamp (SEVC) technique, and 3) application of patch electrodes to central neurons in situ under a dissecting microscope. Using the newly developed treadmill system, the synaptic activity of nonspiking interneurons could be recorded intracellular by a glass microelectrode from an animal walking on it. The SEVC experiments have revealed that the dendritic membrane of nonspiking interneurons possesses three types of depolarization-dependent potassium conductances. Under a dissecting microscope, spontaneous activities of single potassium channels could be recorded by a patch electrode applied to the cell bodies of central neurons in situ exposed by mechanical desheathing of the abdominal ganglion. In invertebrate animals, only a few studies have been carried out so far using the in situ whole-cell recording technique. The results of this study indicate that it is technically possible to apply patch electrodes for whole cell recording to the central neurons in situ in the animal performing behavioral acts so that the synaptic currents associated with specific behavior can be analyzed electrophysiologically.

为了巩固将全细胞记录技术应用于克氏原螯虾（Procambarus clarkii吉拉德）中枢神经系统中执行行为行为的非尖峰中间神经元的实验背景，我将本研究集中在1）微电极和贴片电极可适用的未麻醉的整个动物制备的新发展，2）采用单电极电压钳（SEVC）技术对非尖峰发放的中间神经元的膜电流进行电生理学分析; 3）在解剖显微镜下将贴片电极应用于原位的中枢神经元。利用自行研制的平板运动系统，用玻璃微电极记录了非棘波中间神经元的突触活动，SEVC实验表明非棘波中间神经元树突膜具有三种类型的去极化依赖性钾电导。在解剖显微镜下，将贴片电极应用于经腹神经节机械脱鞘暴露的原位中枢神经元胞体，可记录单个钾通道的自发活动。在无脊椎动物中，迄今为止只有少数研究使用原位全细胞记录技术进行。本研究的结果表明，它是技术上可能的应用贴片电极进行全细胞记录的中枢神经元原位在动物执行行为的行为，使突触电流与特定的行为可以进行电生理分析。