A genetically encoded fluorescent sensor for in vivo imaging of GABA

Jonathan S. Marvin, Yoshiteru Shimoda, Vincent Magloire, Marco Leite, Takashi Kawashima, Thomas P. Jensen, Ilya Kolb, Erika L. Knott, Ondrej Novak, Kaspar Podgorski, Nancy J. Leidenheimer, Dmitri A. Rusakov, Misha B. Ahrens, Dimitri M. Kullmann, Loren L. Looger

Research output: Contribution to journalArticlepeer-review

42 Citations (Scopus)

Abstract

Current techniques for monitoring GABA (γ-aminobutyric acid), the primary inhibitory neurotransmitter in vertebrates, cannot follow transients in intact neural circuits. To develop a GABA sensor, we applied the design principles used to create the fluorescent glutamate receptor iGluSnFR. We used a protein derived from a previously unsequenced Pseudomonas fluorescens strain and performed structure-guided mutagenesis and library screening to obtain intensity-based GABA sensing fluorescence reporter (iGABASnFR) variants. iGABASnFR is genetically encoded, detects GABA release evoked by electric stimulation of afferent fibers in acute brain slices and produces readily detectable fluorescence increases in vivo in mice and zebrafish. We applied iGABASnFR to track mitochondrial GABA content and its modulation by an anticonvulsant, swimming-evoked, GABA-mediated transmission in zebrafish cerebellum, GABA release events during interictal spikes and seizures in awake mice, and found that GABA-mediated tone decreases during isoflurane anesthesia.

Original languageEnglish
Pages (from-to)763-770
Number of pages8
JournalNature Methods
Volume16
Issue number8
DOIs
Publication statusPublished - 2019 Aug 1
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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