Fig. 1: LHA glutamatergic neurons in the hypothalamus are predominantly involved in hindlimb motor control and required for motivation-driven locomotion.

a Left: Schematic of PRV-EGFP injection into the TA (tibialis anterior muscle) and GS (Gastrocnemius muscle). Right: Reconstruction of adult wild-type mouse brain labeled for EGFP after a viral delivery of PRV in the hindlimb muscle. b Reconstruction of PRV-EGFP labelled neurons in the hypothalamus innervating the hindlimb muscles. c The percentage of labelled neurons in the hypothalamus (n = 3). Error bars, SEM. d Three-dimensional heatmap distribution of PRV-labeled cells within the hypothalamus. e The dorsal view (left) and sagittal view (right) of PRV-infected lateral hypothalamus area neurons. f Strategy for activating LHA glutamatergic neurons. g, h Quantification of maximum speed corresponding to optical stimulation in different light intensities and frequencies (n = 6). One-way ANOVA, followed by followed by Dunnett’s multiple comparison. **p < 0.01; ****p < 0.0001^. Error bars, SEM. i Left: Schematic of tetrode recording for LHA neurons and offline spike sorting. Right: Firing rate of one representative neuron in LHA (green line, right axis) plotted with the speed (gray line, left axis) of the mouse. j Distribution of the recorded LHA neurons showing the speed of locomotion based on their relative correlation (r²) with speed. Green bars, speed-correlated neurons; Gray bars, neurons show no significant correlation with the speed. k Top: Raster plot for spike responses of representative neurons to locomotion onset. Bottom: population z-scored firing rate of a subpopulation LHA neurons (green line) and speed (gray line) aligned to the onset of locomotion. Error bars, SEM. l Top: Strategy for fiber photometry recording of LHA Vglut2⁺ neurons and food seeking test in the runway. Bottom: Calcium dynamics of LHA Vglut2⁺ neurons in response to onset of locomotion and consumption. m, Quantification of LHA Vglut2⁺ neurons calcium response to onset of locomotion (top) and food consumption (bottom). n = 7 mice, 3-5 trials per mouse. Student’s t-test (two-tailed, unpaired) was applied. ***p < 0.001; ****p < 0.0001. Error bars, SEM. n Timeline summarizing the experiments of gait analysis, open field and food seeking test with or without depletion of LHA glutamatergic neurons. o Representative stick diagram decomposition of leg movements in mice with or without depletion of LHA glutamatergic neurons. p Quantification of retraction, protraction and height of iliac crest with or without depletion of LHA glutamatergic neurons (n = 6). Student’s t-test (two-tailed, unpaired) was applied. ns not significant. Error bars, SEM. q Top: Trajectory of mice in open-field chambers with or without depletion of LHA glutamatergic neurons. Bottom: Quantification of center time, total distance and entries to center area (n = 6). Student’s t-test (two-tailed, unpaired) was applied. ns, not significant. Error bars, SEM. r, Top: Trajectory of mice in open-field chambers with food pellets placed in the center before and after the depletion of LHA glutamatergic neurons. Bottom: Quantification of entries to center area, total distance, and number of food pellets consumed by mice in open-field chambers with food pellets placed in the center (n = 6). Student’s t-test (two-tailed, unpaired) was applied. *p < 0.05. Error bars, SEM. Source data are provided as a Source data file.