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

为了巩固将全细胞记录技术应用于克氏原螯虾（Procambarus clarkii Girard）中枢神经系统中非尖刺中间神经元进行行为行为的实验背景，我将本研究的重点放在1)一种可应用于微电极和贴片电极的非麻醉全动物制备的新发展；2)利用单电极电压钳（SEVC）技术对非尖峰中间神经元的膜电流进行电生理分析；3)在解剖显微镜下原位膜片电极对中枢神经元的应用。利用新开发的跑步机系统，可以通过行走在其上的动物的玻璃微电极记录细胞内非尖峰中间神经元的突触活动。SEVC实验表明，非尖峰中间神经元的树突膜具有三种类型的去极化依赖钾电导。在解剖显微镜下，将贴片电极贴于经腹神经节机械脱皮暴露的中央神经元细胞体上，可以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了原位全细胞记录技术。本研究的结果表明，在技术上可以应用贴片电极对正在进行行为行为的动物中枢神经元进行全细胞记录，从而对与特定行为相关的突触电流进行电生理学分析。

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.