In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor
In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor
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DOI：
10.21769/bioprotoc.4046
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发表时间：
2021-06-05
期刊：
影响因子：
0.8
通讯作者：
Hirose, Kenzo
中科院分区：
文献类型：
--
作者：
Kitajima, Nami;Takikawa, Kenji;Hirose, Kenzo
腺苷5 '-三磷酸（ATP）作为脑细胞（如神经元和神经胶质细胞）的细胞外信号分子。通过释放ATP的细胞通信涉及正常脑功能所需的一系列过程，异常通信与脑疾病有关。为了研究这些细胞过程的机制，已经开发了各种技术来测量细胞外ATP。为了以高时空分辨率监测细胞外ATP信号传导的动态，我们最近开发了一种混合型ATP光学传感器（ATPOS），该传感器能够在脑中对细胞外ATP动态进行体内荧光成像。ATPOS是通过用小分子荧光染料Cy 3标记ATP结合蛋白芽孢杆菌FoF（1）-ATP合酶e亚基而合成的。将ATPOS注射到活体小鼠的大脑皮层中，能够可视化响应于电刺激的细胞外ATP释放的波状传播。这里描述的协议应该是有用的可视化ATP信号在不同的过程中参与细胞间的通信在大脑中。
Adenosine 5'-triphosphate (ATP) works as an extracellular signaling molecule for cells in the brain, such as neurons and glia. Cellular communication via release of ATP is involved in a range of processes required for normal brain functions, and aberrant communication is associated with brain disorders. To investigate the mechanisms underlying these cellular processes, various techniques have been developed for the measurement of extracellular ATP. To monitor the dynamics of extracellular ATP signaling with high spatiotemporal resolution, we recently developed a hybrid-type ATP optical sensor (ATPOS) that enables in vivo fluorescence imaging of extracellular ATP dynamics in the brain. ATPOS is synthesized by labeling an ATP-binding protein, Bacillus FoF(1)-ATP synthase e subunit, with a smallmolecular fluorescent dye Cy3. Injection of ATPOS into the cerebral cortex of living mice enables visualization of the wave-like propagation of extracellular ATP release in response to electrical stimulation. The protocol described here should be useful for visualizing ATP signaling in diverse processes involved in intercellular communication in the brain.