Fig. 6: In vivo l-lactate dynamics during neural activity. | Nature Communications

Fig. 6: In vivo l-lactate dynamics during neural activity.

From: A red fluorescent genetically encoded biosensor for in vivo imaging of extracellular l-lactate dynamics

Fig. 6

a The setup for recording of R-eLACCO2.1 and locomotion in an awake mouse under air-puff stimulation (20 s of 4 Hz) to multiple vibrissae. ΔF/F map (bottom) of the same field of view averaged for 60–80 s after a vibrissa stimulation onset. Scale bars, 100 μm. b ΔF/F traces of R-eLACCO2.1 under air-puff stimulation and spontaneous locomotion after 30 s rest in an example mouse (mean ± s.d. of the 30 cells indicated in panel a). c Averaged ΔF/F traces of hSyn-R-eLACCO2.1 (n = 107 locomotion events, n = 48 vibrissae stimulations in 5 mice) and hSyn-R-deLACCOctrl (n = 59 locomotion events, n = 53 vibrissae stimulations in 3 mice) under spontaneous locomotion following 30-s rest (upper) and air-puff stimulation (bottom). Mean ± s.e.m. Two-tailed Welch’s unpaired t-test. **p = 0.0001 (locomotion events), *p < 0.0001 (vibrissae stimulation). d Schematic illustration of simultaneous imaging of l-lactate and Ca2+ in awake mice under AO2P. Mice co-expressing R-eLACCO2.1 (red, excited at 1,030 nm) and GCaMP6f (green, excited at 1,030 nm) in vS1 neurons were i.v. injected with Cy5.5 dye (vasculature, cyan, excited at 950 nm). Scale bar, 100 µm. e Verification of R-eLACCO2.1 and GCaMP6f expression in one-photon widefield imaging (left top, scale bar 100 µm) and AO2P imaging (left bottom and right, scale bars 20 µm). f Representative ΔF/F traces of neurons co-expressing R-eLACCO2.1 and GCaMP6f in an awake mouse identified on a maximum intensity projection. n = 20 neurons and 46 Ca2+ events after 30 seconds of quiescence from one mouse. Mean ± s.d. Scale bar, 20 µm. g ΔF/F traces of neurons co-expressing GCaMP6f and R-eLACCO2.1 (left, n = 80 neurons and 247 Ca2+ events after 30 seconds of quiescence from 4 mice) or R-deLACCOctrl (right, n = 188 neurons and 478 Ca2+ events after 30 seconds of quiescence from 3 mice) in awake mice. Mean ± s.e.m. The increases in fluorescence observed for both R-eLACCO2.1 and R-deLACCOctrl at t = 0 are attributable to bleedthrough from the GCaMP6f signal. Source data are provided as a Source Data file.

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