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High-Throughput Development of a Hybrid-Type Fluorescent Glutamate Sensor for Analysis of Synaptic Transmission by Kenji Takikawa; Daisuke Asanuma; Shigeyuki Namiki; Hirokazu Sakamoto; Tetsuro Ariyoshi; Naoya Kimpara; Kenzo Hirose is a Chemistry article available to read on EtoBox.
What is High-Throughput Development of a Hybrid-Type Fluorescent Glutamate Sensor for Analysis of Synaptic Transmission about?
Fluorescent sensors are powerful tools for visualizing cellular molecular dynamics. We present a high-throughput screening system, designated hybrid-type fluorescence indicator development (HyFInD), to identify optimal positionspecific fluorophore labeling in hybrid-type sensors consisting of combinations of ligand-binding protein mutants with small molecular fluorophores. We screened sensors for glutamate among hybrid molecules obtained by the reaction of four cysteine-reactive fluorescence probes with a set of cysteinescanning mutants of the 274 amino acid S1S2 domain of AMPA-type glutamate receptor GluA2 subunit. HyFInD identified a glutamate-responsive probe (enhanced glutamate optical sensor: eEOS) with a dynamic range > 2400 %, good photostability, and high selectivity. When eEOS was specifically tethered to neuronal surfaces, it reliably visualized the spatiotemporal dynamics of glutamate release at single synapses, revealing synapse-to-synapse heterogeneity of short-term plasticity.
Who reads High-Throughput Development of a Hybrid-Type Fluorescent Glutamate Sensor for Analysis of Synaptic Transmission?
It is typically read by researchers, students, and practitioners in Chemistry.
- Author
- Kenji Takikawa; Daisuke Asanuma; Shigeyuki Namiki; Hirokazu Sakamoto; Tetsuro Ariyoshi; Naoya Kimpara; Kenzo Hirose
- Publisher
- Wiley
- Published
- 2014
- Language
- EN
- Field
- Chemistry (Physical Sciences)