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Calcium-independent biosensors for multiplexed in vivo imaging of extracellular lactate dynamics

Liu, J.-J.; Mizuno, H.; Than-Trong, E.; Machler, P.; Kubotera, H.; Barbay, T.; Wang, C.-Y.; To, Q.-L.; Kamijo, Y.; Nishimura, T.; Boisvert, M.; Paquet, M.-E.; Drobizhev, M.; Cragg, S. J.; Nagai, J.; Kleinfeld, D.; Charpak, S.; NASU, Y.

2026-10-05 molecular biology
10.64898/2026.10.02.756141 bioRxiv
Show abstract

Extracellular L-lactate reports cellular metabolism, but resolving its dynamics alongside intracellular metabolism and neuronal activity remains challenging. Here we develop eLACCO3 and R-eLACCO3, green and red cell-surface lactate biosensors with Ca2+-independent responses and faster response and recovery kinetics in cultured cells. Selected variants provide larger fluorescence responses than their predecessors in brain cells, slices, and living mice and improve detection of glucose-evoked extracellular lactate oscillations. Combining R-eLACCO3 with GCaMP6f enabled imaging of extracellular L-lactate simultaneously with odor-evoked activity in anesthetized mice and cortical activity in awake mice. These biosensors enable multiplexed measurements of extracellular lactate dynamics in relation to intracellular metabolism and neuronal activity.

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