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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.

研究大量神经活动的一种方法是使用荧光电压敏感染料同时跟踪许多神经元的神经活动。电压敏感染料首先应用于嗅球的神经元。然后用紫外线照射灯泡，并通过显微镜拍摄暴露的嗅球的视频图像。紫外光刺激电压敏感染料，当它所在的细胞去极化时发出光。通过使用计算机从存在气味时获得的图像中减去背景图像。我们在家蚕蛾脑中进行了活体光学记录，以研究神经元群体对传递给动物的气味剂的反应。成虫雄蛾在羽化后2-4天内使用。在4℃冷却约30 min麻醉后，在大脑上方打开头囊，切除头部大部分肌肉。用细镊子将脑去鞘并用电压敏感染料RH 414（分子探针）染色。将RH 414染料以3 mg/ml的浓度溶解在生理盐水中。用盐水洗去过量的染料。将信号叠加在AL的亮场图像上，以将响应与AL结构直接相关。气味诱发特定的时空兴奋模式在AL。特定的合奏活动肾小球可能代表气味的组合方式。气味混合物诱发的模式是单一气味反应的组合。气味不仅表现为空间活动模式，而且表现为随时间演变的模式。