Laser trapping of menbrane protein particles in neurons

激光捕获神经元中的膜蛋白颗粒

• 批准号: 06808070
• 负责人: TATSUMI Hitoshi
• 金额: $ 1.15万
• 依托单位: TOKYO MEDICAL AND DENTAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06808070/
• 关键词: Nano Meter; Neuron; Growth cone; synaps; Laser trap; membrane; protein; Nano-meter; molecule; 微分干渉顕微鏡; ビデオ画像処理; ナノメートル計測; 光ピンセット; レーザー光; 光学技術; 神経生理学; 金コロイド粒子

Neurons protrude growth cones during development and also during axon sprouting after damage. Evoked release of acetylcholine (ACh) from the growth cones of central cholinergic neurons in culture was monitored using whole-cell patch clamp recording from ACh-receptor-rich neuron placed close to the growth cones as a sensor of ACh release. Simultaneous video-enhanced DIC imaging of growth cones showed contacts between growth cones and the sensor neuron after ACh release. Adhesion was also observed between growth cones and latex beads, when growth cones were exposed to high potassium solution. After beads attached on the growth cone membrane, 1 pN of laser optical pressure was required for moving the beads about 0z1 to 1 mum on the growth cone surface. The changes in the surface viscosity of the growth cone membrane was estimated by analyzing the video images of laser trapped beads on it.

神经元在发育过程中以及损伤后轴突发芽过程中突出生长锥。采用全细胞膜片钳技术，以生长锥附近富含乙酰胆碱（ACh）受体的神经元作为乙酰胆碱（ACh）释放的传感器，监测培养的中枢胆碱能神经元生长锥的乙酰胆碱（ACh）释放。同时视频增强DIC成像的生长锥显示ACh释放后生长锥和传感器神经元之间的联系。当生长锥暴露于高钾溶液中时，生长锥和乳胶珠之间也观察到粘附。在珠粒附着在生长锥膜上之后，需要1pN的激光光学压力来使珠粒在生长锥表面上移动约0 μ m至1 μ m。通过分析激光捕获珠在生长锥膜上的视频图像，估计了生长锥膜表面粘度的变化。

项目成果

辰巳仁史: "細胞内カルシウム実験プロトコール" 羊士社, 201〜 (1996)

辰巳仁：《细胞内钙实验方案》Yoshisha，201~（1996）

Tatsumi: Protocol for cellular Calcium research, Imaging microscopy. Yodo-sha Co.Ltd., 47-58 (1996)

Tatsumi：细胞钙研究方案，成像显微镜。

辰己仁史: "実験医学別冊 脳・神経プロトコール、光ピンセット" 羊土社, 204-206

Hitoshi Tatsumi：“实验医学特刊：大脑/神经协议，光镊” Yodosha，204-206

Tatsumi,H and Y.Katayama: "Analysis of Ca^<2+> homeostasis in newons dissociated from rat nucleus basalis" Neuroscience Research. (accepted).

Tatsumi,H 和 Y.Katayama：“从大鼠基底核分离的牛顿中 Ca^<2> 稳态的分析”神经科学研究。

Tatsumi H.Tsuji S.et al.: "Synthesis,storage and release of acehylcholine at and from growth cones・・・" Neuroscience Lett.202. 25-28 (1995)

Tatsumi H.Tsuji S.et al.：“乙酰胆碱在生长锥中的合成、储存和释放......”《神经科学快报》25-28 (1995)。