Establishment of evolutional microscopy on neurons with ultra-fast two-photon microscope

超快双光子显微镜神经元进化显微镜的建立

• 批准号: 23300121
• 负责人: INOUE Takafumi
• 金额: $ 12.81万
• 依托单位: Waseda University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23300121/
• 关键词: 分子動態; タンパク質; 神経細胞; シナプス; 活動電位; 膜電位計測; 樹状突起; スパイン; ２光子励起顕微鏡; 蛍光相関分光法; 拡散; 2光子励起顕微鏡; 分子・細胞神経科学; タンパク質の動態; 分子動態; タンパク質; 神経細胞; シナプス; 活動電位; 膜電位計測; 樹状突起; スパイン; ２光子励起顕微鏡; 蛍光相関分光法; 拡散; 2光子励起顕微鏡; 分子・細胞神経科学; タンパク質の動態

This research project aimed to measure dynamics of protein molecules rapidly moving in the cytosol of neuronal dendrites at multiple points simultaneously by means of a newly developed two-photon microscope. I succeeded in recording at multiple points at more than 10 kHz of time resolution from cytosol of primary cultured neurons derived from mouse, in which GFP or GFP-fused CamKII proteins were expressed, and in analyzing by FCS. I was also successful in recording action potentials from multiple cultured neurons, which enabled to analyze synaptic connections between neuron pairs and also direction of synaptic connections.

该研究项目旨在通过新开发的两光子显微镜同时在多个点上迅速在神经元树突的细胞质中迅速移动蛋白质分子的动力学。我成功地以超过10 kHz的时间分辨率记录了来自小鼠的原代神经元的细胞质，其中表达了GFP或GFP融合的CAMKII蛋白，并通过FCS进行分析。我还成功地记录了来自多个培养神经元的动作电位，这使得能够分析神经元对之间的突触连接以及突触连接的方向。