Optical Recording and Analysis of Neural Activities in Odor response of Larva Silkworm Moth

蚕蛾幼虫气味反应神经活动的光学记录与分析

基本信息

批准号：
14350437
负责人：
OHTAKE Hisao
金额：
$ 9.22万
依托单位：
Osaka University (2003-2004)Hiroshima University (2002)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14350437/
关键词：
Silkworm moth Olfactory Mulberry tree Electrophysiological experiments

项目摘要

One approach to the study of massed neural activity has been to use fluorescent voltage-sensitive dyes to track the neural activity of many neurons at once. The voltage-sensitive dye is first applied to the neurons of the olfactory bulb. The bulb is then illuminated with ultraviolet light, and video images of the exposed olfactory bulb are taken through a microscope. The UV light stimulates the voltage-sensitive dye to give off light when the cell in which it is located depolarizes. By using a computer to subtract the background image from the image obtained when the odor is present. We made optical recordings in vivo in the silkworm moth brain to investigate neuronal population responses to odorants delivered to the animal. Adult male silkworm moths were used within 2-4 days after eclosion. After anesthesia by cooling at 4℃ for about 30 min, the head capsule was opened above the brain, and most of the muscles in the head were removed. The brain was desheathed by fine tweezers and stained with a voltage-sensitive dye RH414 (Molecular probe). The RH414 dye was dissolved in a physiological saline at a concentration of 3 mg/ml. Excess dye was washed off with the saline. The signals were superimposed on the bright-field image of AL to correlate the responses directly with the AL structure. Odors evoked specific spatio-temporal excitation patterns in the AL. Specific ensembles of active glomeruli are likely to represent odors in a combinatorial manner. Odor blends evoked patterns that were combinations of the single odorant responses. Odors are not only represented as spatial activity patterns, but as patterns that evolve over time.

研究大量神经元活性的一种方法是使用荧光电压敏感染料一次跟踪许多神经元的神经元活性。电压敏感的染料首先应用于嗅球的神经元。然后用紫外线照亮灯泡，并通过显微镜拍摄裸露的嗅球的视频图像。紫外线光刺激对电压敏感的染料，当其位于其位于其的细胞时散发光。通过使用计算机从出现气味时获得的图像中减去背景图像。我们在蚕蛾大脑的体内进行了光学记录，以研究对动物传递的气味的神经元种群的反应。成年雄性蚕飞蛾在ECLOSIN后2-4天内使用。通过在4℃冷却大约30分钟的麻醉后，将头囊打开大脑上方，并去除头部大部分肌肉。大脑被细的镊子消失，并用电压敏感的染料RH414（分子探针）染色。将RH414染料溶解在物理盐水中，浓度为3 mg/ml。多余的染料被盐水冲洗掉。信号叠加在AL的明亮场图像上，以将响应直接与AL结构相关联。气味引起了AL中的特定时空兴奋模式。活性肾小球的特定集合可能以组合方式代表气味。气味混合了诱发的模式，这些模式是单个气味反应的组合。气味不仅表示为空间活动模式，而且是随着时间的流逝而发展的模式。