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An optimized fluorescent probe for visualizing glutamate neurotransmission.
Nat Methods ; 10(2): 162-70, 2013 Feb.
Article em En | MEDLINE | ID: mdl-23314171
We describe an intensity-based glutamate-sensing fluorescent reporter (iGluSnFR) with signal-to-noise ratio and kinetics appropriate for in vivo imaging. We engineered iGluSnFR in vitro to maximize its fluorescence change, and we validated its utility for visualizing glutamate release by neurons and astrocytes in increasingly intact neurological systems. In hippocampal culture, iGluSnFR detected single field stimulus-evoked glutamate release events. In pyramidal neurons in acute brain slices, glutamate uncaging at single spines showed that iGluSnFR responds robustly and specifically to glutamate in situ, and responses correlate with voltage changes. In mouse retina, iGluSnFR-expressing neurons showed intact light-evoked excitatory currents, and the sensor revealed tonic glutamate signaling in response to light stimuli. In worms, glutamate signals preceded and predicted postsynaptic calcium transients. In zebrafish, iGluSnFR revealed spatial organization of direction-selective synaptic activity in the optic tectum. Finally, in mouse forelimb motor cortex, iGluSnFR expression in layer V pyramidal neurons revealed task-dependent single-spine activity during running.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes de Fusão / Transmissão Sináptica / Ácido Glutâmico / Proteínas de Escherichia coli / Proteínas de Fluorescência Verde / Corantes Fluorescentes Limite: Animals Idioma: En Ano de publicação: 2013 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes de Fusão / Transmissão Sináptica / Ácido Glutâmico / Proteínas de Escherichia coli / Proteínas de Fluorescência Verde / Corantes Fluorescentes Limite: Animals Idioma: En Ano de publicação: 2013 Tipo de documento: Article