FORTIS: a live-cell assay to monitor AMPA receptors using pH-sensitive fluorescence tags

• Main Authors: María Calleja-Felipe, Magdalena Natalia Wojtas, Marta Diaz-González, Dalila Ciceri, Raúl Escribano, Alberto Ouro, Miguel Morales, Shira Knafo
• Format: Article
• Language: English
• Published: Nature Publishing Group 2021-05-01
• Series: Translational Psychiatry
• Online Access: https://doi.org/10.1038/s41398-021-01457-w

Abstract 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.