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

Development of an optical recording system for neuronal network activities of invertebrates

无脊椎动物神经网络活动光学记录系统的开发

基本信息

批准号：
07554071
负责人：
TAKAHATA Masakazu
金额：
$ 1.86万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07554071/
关键词：
invertebrate crayfish optical recording system dendrite synaptic activity calcium fluorescence indicator image processing central neuron 蛍光細胞内染色 LDS細胞画像解析入力シナプス

项目摘要

An optical recording system that can be used in electrophysiological experiments using the whole animal preparation of invertebrates has been developed to visualize the change in intracellular calcium concentration associated with the synaptic activity. The system consists of an epifluorescence device for exciting Fura-2 that is iontophoretically injected into a cell, a dissecting microscope with the working distance of 187mm, a Peltier-cooled CCD video camera, and a personal computer with newly developed programs for image analysis. To test the performance of this recording system, nerve cells in the terminal abdominal ganglion of crayfish were loaded with the dye and observed from the dorsal side with the CCD exposure time of 16sec. The whole cell structure having the soma, dendrites and axon could be visualized in situ together with the electrode tip under the dissecting microscope by either 380nm or 340nm excitation. When the sensory nerve bundle was stimulated repeatedly at 1Hz during the exposure time, a decrease in the fluorescence of the dye to excitation at 380nm was detected in a specific dendritic region of the cell. However, any significant change in the ratio of fluorescence by 380nm to that by 340nm excitation during synaptic activation could not be detected probably due to the relatively weak strength of the latter. It is concluded that although the sensitivity and time resolution are not enough to quantify the dynamics of intracellular calcium ions, the presently developed system can be used to localize the functional synaptic region within a cell by visualizing the change in intracellular calcium concentration due to synaptic activity together with the whole cell structure.

一个光学记录系统，可用于电生理实验中使用的整个动物制备的无脊椎动物已经开发出可视化的细胞内钙离子浓度的变化与突触活动。该系统包括一个用于激发Fura-2的荧光装置，一个工作距离为187 mm的解剖显微镜，一个Peltier冷却的CCD摄像机，和一个带有新开发的图像分析程序的个人计算机。为了测试该记录系统的性能，在螯虾的终末腹神经节的神经细胞负载染料，并从背侧观察与CCD曝光时间为16秒。在解剖显微镜下，通过380 nm或340 nm的激发，可以与电极尖端一起原位观察到包括索马、树突和轴突的整个细胞结构。当感觉神经束在暴露时间期间以1Hz重复刺激时，在细胞的特定树突区域中检测到染料对380 nm激发的荧光的减少。然而，在突触激活过程中，380 nm与340 nm激发的荧光的比率的任何显著变化不能被检测到，可能是由于后者的强度相对较弱。它的结论是，虽然灵敏度和时间分辨率是不够的，以量化细胞内钙离子的动态，目前开发的系统可以用来本地化的功能突触区域内的细胞内的可视化细胞内钙浓度的变化，由于突触活动与整个细胞结构。