MacDonald, RB; Kashikar, ND; Lagnado, L; Harris, WA; (2017) A Novel Tool to Measure Extracellular Glutamate in the Zebrafish Nervous System In Vivo. Zebrafish , 14 (3) pp. 284-286. 10.1089/zeb.2016.1385.

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Abstract

Glutamate is the major excitatory neurotransmitter in the brain. Its release and eventual recycling are key to rapid sustained neural activity. We have paired the gfap promoter region with the glutamate reporter molecule, iGluSnFR, to drive expression in glial cells throughout the nervous system. Tg(gfap:iGluSnFR) is expressed on the glial membrane of Müller glia cells in the retina, which rapidly respond to stimulation and the release of extracellular glutamate. As glial cells are associated with most, if not all, synapses, Tg(gfap:iGluSnFR) is a novel and exciting tool to measure neuronal activity and extracellular glutamate dynamics in many regions of the nervous system. Glutamate is the major excitatory neurotransmitter in the brain. Its release and eventual recycling are key to rapid sustained neural activity.1 Glial cells play a key role in the uptake and recycling of glutamate from the synaptic cleft. iGluSnFR has been used to study synaptic activity by measuring glutamate dynamics in the zebrafish nervous system.2,3 Previous work has also used iGluSnFR in glial cells; however, this was done transiently in the mouse using viral vectors.2,4 As such, we designed a transgene to stably express iGluSnFR in the glial cells of the zebrafish nervous system. We report a novel transgenic zebrafish, Tg(gfap:iGluSnFR), that displays the glutamate-sensitive fluorescent reporter iGluSnFR specifically on the membrane of glial cells (Figure 1A–C). This molecule is expressed on the glial membrane in many brain regions and rapidly responds to stimulation and the release of extracellular glutamate (Figure 1D–F, Supplementary Data; Supplementary Data are available online at www.liebertpub.com/zeb). Thus, pairing the sensitivity of iGluSnFR and optical transparency of the zebrafish provides a powerful tool for understanding glutamate dynamics in neural tissues in vivo.

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