Background: Neurobiologic abnormalities in dorsolateral prefrontal cortex (DLPFC) are believed to be involved in the pathophysiology of major depressive disorder (MDD). Although MDD commonly emerges during childhood and adolescence, to out-knowledge, no prior study has examined the DLPFC in pediatric patients with MDD. Methods: In this study, choline compounds (Cho), N-acetylaspartate (NAA), and creatine/phosphocreatine (Cr) were measured in left and right DLPFC using a multislice proton magnetic resonance spectroscopic imaging sequence with validated phantom replacement methodology in 11 treatment-naive MDD patients, 10-16 years of age, and 11 case-matched healthy control subjects. Results: A significant increase in Cho was observed in left but not right DLPFC in MDD patients versus control subjects (32.5% higher). No significant differences in NAA or Cr were observed between case-control pairs. Conclusions: These results provide new evidence of localized functional neurochemical marker alterationsi in left DLPFC in pediatric MDD. Our results must be considered preliminary, however, given the small sample size.