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Alterations in the mirror neuron system (MNS), crucial for interpreting and imitating actions, may underlie these ASD-associated differences in motor imitation. High-density diffuse optical tomography (HD-DOT) overcomes logistical challenges of functional magnetic resonance imaging to enable identification of neural substrates of naturalistic motor imitation. We aim to investigate brain function underlying motor observation and imitation in autistic and non-autistic adults. We hypothesize that HD-DOT will reveal greater activation in regions associated with the MNS during motor imitation than during motor observation, and that MNS activity will negatively correlate with autistic traits and motor fidelity. We imaged brain function using HD-DOT in N = 100 participants (19 ASD and 81 non-autistic individuals) as they engaged in observing or imitating a sequence of arm movements. Additionally, during imitation, participant movements were simultaneously recorded with 3D cameras for computerized assessment of motor imitation (CAMI). Cortical responses were estimated using general linear models, and multiple regression was used to test for associations with autistic traits, assessed via the Social Responsiveness Scale-2 (SRS-2), and imitation fidelity, assessed via CAMI. Both observing and imitating motor movements elicited significant activations in higher-order visual and MNS regions, including the inferior parietal lobule, superior temporal gyrus, and inferior frontal gyrus. Imitation additionally exhibited greater activation in the superior parietal lobule, primary motor cortex, and supplementary motor area. Notably, the right temporal\u2013parietal junction exhibited activation during observation but not during imitation. Higher presence of autistic traits was associated with increased activation during motor observation in the right superior parietal lobule. No significant associations between brain activation and CAMI scores were observed. Our findings provide robust evidence of shared and task-specific cortical responses underlying motor observation and imitation, emphasizing the differential engagement of MNS regions during motor observation and imitation.<\/jats:p>","DOI":"10.1162\/imag.a.153","type":"journal-article","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T20:26:43Z","timestamp":1756844803000},"update-policy":"https:\/\/doi.org\/10.1162\/mitpressjournals.corrections.policy","source":"Crossref","is-referenced-by-count":2,"title":["Mapping brain function underlying naturalistic motor observation and imitation using high-density diffuse optical tomography"],"prefix":"10.1162","volume":"3","author":[{"given":"Dalin","family":"Yang","sequence":"first","affiliation":[{"name":"Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, MO, United States"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tessa 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