Extended Data Fig. 4: High-frequency STN DBS excites the presynaptic afferent terminals in the STN of normal mice.

a, Schematic illustration of the strategy to express GCaMP7f in the afferent terminals in the STN and tdTomato in STN neurons of Vglut2-cre mice. b, Immunofluorescence images to show the expression of GCaMP7f in the cortex and GPe, and GCaMP7f and tdTomato in the STN. Scale bars: 200 μm, except in the small insert of Cortex (M1/M2), which is 500 μm. c, Illustrations of fiber photometry recordings of presynaptic Ca²⁺ levels in the STN during STN DBS using GCaMP7f expressed in afferent terminals and tdTomato expressed in STN neurons. tdTomato serves as a control fluorescent protein and a marker for STN to guide the probe implantation. d, Representative emission spectra of GCaMP7f expressed by presynaptic axonal terminals and tdTomato expressed by STN neurons before and during DBS stimulation. e, f, Time-lapsed ratio of GCaMP7f/tdTomato fluorescence (normalized to the baseline before the first DBS stimulation) to show the DBS-induced changes in presynaptic Ca²⁺ levels in afferent terminals in the STN at different stimulation intensities (e) and at the same intensity (f). g, Summary of DBS-induced increases in presynaptic Ca²⁺ levels in afferent terminals in the STN at different stimulation intensities. * p < 0.05, ** p < 0.01 (100 µA vs. 150 µA, p = 0.0198; 150 µA vs. 200 µA, p = 0.0081), Repeated one-way ANOVA followed by Dunnett’s multiple comparisons test. All data are plotted as mean ± SEM. n = 7 mice for each group, including both males and females.

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