To investigate the effects of omega-3 fatty acid deficiency on phosphatidylinositol signaling in brain, myoinositol (mI) concentrations were determined in the prefrontal cortex (PFC) of omega-3 fatty acid deficient rats by in vivo proton magnetic resonance spectroscopy ( 1 H-MRS). To generate graded deficits in PFC docosahexaenoic acid (22:6n-3) (DHA) composition, perinatal and postweaning a-linolenic acid (18:3n-3) (ALA) deficiency models were used. Adult male rats were scanned in a 7T Bruker Biospec system and a 1 H-MRS spectrum acquired from the bilateral medial PFC. Rats were then challenged with SKF83959, a selective agonist at phosphoinositide (PI)-coupled dopamine D 1 receptors. Postmortem PFC fatty acid composition was determined by gas chromatography. Relative to controls, PFC DHA composition was significantly reduced in adult postweaning (227%) and perinatal (265%) ALA-deficiency groups. Basal PFC mI concentrations were significantly reduced in the perinatal deficiency group (221%, P 5 0.001), but not in the postweaning deficiency group (21%, P 5 0.86). Among all rats, DHA composition was positively correlated with mI concentrations and the mI/creatine (Cr) ratio. SKF83959 challenge significantly increased mI concentrations only in the perinatal deficiency group (116%, P 5 0.02). These data demonstrate that perinatal deficits in cortical DHA accrual significantly and selectively reduce mI concentrations and augment receptor-generated mI synthesis.-McNamara, R. K., J. Able, R. Jandacek, T. Rider, P. Tso, and D. M. Lindquist. Perinatal n-3 fatty acid deficiency selectively reduces myo-inositol levels in the adult rat PFC: an in vivo 1 H-MRS study. J. Lipid Res. 2009. 50: 405-411.