FORTIS：一种使用 pH 敏感荧光标签监测 AMPA 受体的活细胞检测法。
FORTIS: a live-cell assay to monitor AMPA receptors using pH-sensitive fluorescence tags.
机构信息
Department of Physiology and Cell Biology, Faculty of Health Sciences, The National Institute for Biotechnology in the Negev, and The Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Instituto Biofisika (UPV/EHU, CSIC), University of the Basque Country, Leioa, E-48940, Spain.
出版信息
Transl Psychiatry. 2021 May 27;11(1):324. doi: 10.1038/s41398-021-01457-w.
The real-time live fluorescent monitoring of surface AMPA receptors (AMPARs) could open new opportunities for drug discovery and phenotypic screening concerning neuropsychiatric disorders. We have developed FORTIS, a tool based on pH sensitivity capable of detecting subtle changes in surface AMPARs at a neuronal population level. The expression of SEP-GluA1 or pHuji-GluA1 recombinant AMPAR subunits in mammalian neurons cultured in 96-well plates enables surface AMPARs to be monitored with a microplate reader. Thus, FORTIS can register rapid changes in surface AMPARs induced by drugs or genetic modifications without having to rely on conventional electrophysiology or imaging. By combining FORTIS with pharmacological manipulations, basal surface AMPARs, and plasticity-like changes can be monitored. We expect that employing FORTIS to screen for changes in surface AMPARs will accelerate both neuroscience research and drug discovery.
实时荧光监测表面 AMPA 受体 (AMPARs) 可以为神经精神疾病的药物发现和表型筛选开辟新的机会。我们开发了 FORTIS，这是一种基于 pH 敏感性的工具，能够在神经元群体水平上检测到表面 AMPARs 的细微变化。在 96 孔板中培养的哺乳动物神经元中表达 SEP-GluA1 或 pHuji-GluA1 重组 AMPAR 亚基，可以用微孔板读数器监测表面 AMPARs。因此，FORTIS 可以在不依赖传统电生理学或成像的情况下，记录药物或基因修饰引起的表面 AMPARs 的快速变化。通过将 FORTIS 与药理学操作相结合，可以监测基础表面 AMPARs 和类似可塑性的变化。我们预计，利用 FORTIS 筛选表面 AMPARs 的变化将加速神经科学研究和药物发现。
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