This study compared the effects of chemically synthesized psilocybin (PSIL) and psychedelic mushroom extract (PME) on the head twitch response (HTR), synaptic proteins, and metabolomic profiles in male C57Bl/6j mice. The HTR measurements showed similar effects of PSIL and PME over 20 minutes. Western blot analysis revealed that both treatments significantly increased the levels of synaptic proteins GAP43 and synaptophysin in the frontal cortex and hippocampus, and SV2A in the amygdala. PME also increased GAP43 in the hippocampus and SV2A in the hippocampus, while PSIL increased GAP43 in the amygdala. Metabolomic analyses using capillary electrophoresis - Fourier transform mass spectrometry (CE-FTMS) showed a clear separation between PME and vehicle groups, with purines showing a progressive decline from vehicle to PSIL to PME. The study suggests that PME has a more potent and prolonged effect on synaptic plasticity compared to PSIL, and that PME may have a different therapeutic mechanism compared to PSIL. Further research is needed to confirm these findings and identify the molecules responsible for the enhanced effects of PME.This study compared the effects of chemically synthesized psilocybin (PSIL) and psychedelic mushroom extract (PME) on the head twitch response (HTR), synaptic proteins, and metabolomic profiles in male C57Bl/6j mice. The HTR measurements showed similar effects of PSIL and PME over 20 minutes. Western blot analysis revealed that both treatments significantly increased the levels of synaptic proteins GAP43 and synaptophysin in the frontal cortex and hippocampus, and SV2A in the amygdala. PME also increased GAP43 in the hippocampus and SV2A in the hippocampus, while PSIL increased GAP43 in the amygdala. Metabolomic analyses using capillary electrophoresis - Fourier transform mass spectrometry (CE-FTMS) showed a clear separation between PME and vehicle groups, with purines showing a progressive decline from vehicle to PSIL to PME. The study suggests that PME has a more potent and prolonged effect on synaptic plasticity compared to PSIL, and that PME may have a different therapeutic mechanism compared to PSIL. Further research is needed to confirm these findings and identify the molecules responsible for the enhanced effects of PME.