Background: The lateral hypothalamus (LH), known to play roles in modulation of reward, avoidance, and preference, projects to the associated brain regions such as the ventral tegmental area (VTA), lateral habenula (LHb), and rostromedial tegmental nucleus (RMTg). Previous studies have established that LH neurons projecting to LHb influence cocaine-induced behavior and dopamine release. However, the specific neuronal populations and the peptides involved in cocaine-induced behavioral effects have not been elucidated.Purpose: To address this, our study aimed to investigate the roles of LH neuropeptides including MCH, orexin/hypocretin, GABA, and glutamate in cocaine-enhanced locomotor behaviors and 50-kHz ultrasonic vocalizations (USVs).Materials and Methods: Using antagonists, we selectively inhibited the neuropeptides from LH neurons and assessed cocaine-induced locomotion. We implanted guide cannula into LHb, injected the antagonists of MCH, orexin, GABA, and glutamate prior to cocaine injection and monitored locomotor behaviors up to 1.5 hr after cocaine injection.Results: Antagonists of glutamate or orexin blocked cocaine-induced locomotion and 50-kHz USVs. The MCH antagonist increased cocaine-induced locomotion and 50-kHz USVs.Conclusion: Our findings highlight a significant role for orexin, glutamate and MCH, suggesting these neurons mediate the psychomotor effects induced by cocaine but play distinct roles in the process.This work was supported by the Korea Medical Device Development Fund grant funded by the Korea government(the Ministry of Science and ICT, the Ministry of Trade, Industry and Energy, the Ministry of Health&Welfare, the Ministry of Food and Drug Safety) RS-2023-00253560 and National Research Foundation of Korea(NRF) Grants funded by the Korea government(MSIT) RS-2023-00262398.