Six subjects with treatment resistant major depressive disorder (MDD) were scanned with resting state functional connectivity (rsfMRI)and GABA/Glx editing protocol. rsFMRI between left dorsolateral prefrontal cortex (lDLPFC) and left anterior insula (lAI), two nodes of physiological importance in MDD, inversely correlated with Glx at DLPFC. No such association was observed between rsfMRI and GABA. lDLPFC-lAI rsfMRI, lDLPFC GABA or Glx levels did not correlate with disease severity. The results suggest a collective role of lDLPFC and lAI in MDD via a glutamatergic mechanism.