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GluTrooper: a novel reporter mouse line for whole-brain imaging of glutamate dynamics

Calandra, G. M.; Varga, D. P.; Feyen, P. L. C.; Gubinelli, F.; Wefers, B.; Makra, P.; Shrouder, J. J.; Cattaneo, A.; Gökce, S.; Wurst, W.; Liesz, A.; Herms, J.; Burbulla, L.; Filser, S.; Plesnila, N.

2025-11-12 neuroscience
10.1101/2025.11.11.686940 bioRxiv
Show abstract

Glutamate is the primary excitatory neurotransmitter in the mammalian brain. However, tools to image glutamate dynamics in the whole brain with high spatial and temporal resolution are lacking. Therefore, we developed GluTrooper, a novel mouse line engineered for inducible and long-lasting expression of the genetically encoded glutamate sensor iGluSnFR3. GluTrooper mice crossed with Emx1-Cre lines demonstrated uniform and stable sensor expression in excitatory neurons of the cortex, hippocampus, and olfactory bulb. iGluSnFR3 expression remained stable for at least 12 months, enabling longitudinal observations of glutamate dynamics over extended periods. Using multimodal imaging in awake mice, we demonstrated the versatility of GluTrooper across multiple spatial scales: from mesoscale widefield cortical imaging to cellular resolution with two-photon microscopy. Moreover, during cortical spreading depolarization, bilateral whole-brain glutamate dynamics and contralateral cortical disinhibition were detected with high fidelity. Accordingly, the GluTrooper may open new avenues for the better understanding of glutamatergic neurotransmission in the mammalian brain.

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