Long-chain PUFA (LC-PUFA) are important for fetal and neonatal brain development. However, their accretion in the brain is compromised during maternal protein restriction. Hence, we investigated the effect of maternal supplementation with n-3 DHA plus n-6 arachidonic acid (ARA) at a low protein level (9 %) on offspring brain fatty acid accretion using Wistar rats (nine rats per group) randomly fed a control (C), a low-protein (LP) or a low-protein DHA + ARA-supplemented (LPS) diet during gestation and lactation. At birth, pups from the LPS group had the highest brain DHA and n-3 fatty acid levels (P = 0·001), whereas pups from the LP group had the highest MUFA (P = 0·05) but the lowest DHA and total n-3 PUFA levels (P = 0·000). During lactation, pups from the LPS group accrued significantly more α-linolenic acid (P = 0·003), EPA (P = 0·02) and DHA (P = 0·000) in brain lipids than pups from the LP group, whereas brain lipids of pups from the LP group had markedly increased levels of the n-3 deficiency marker docosapentaenoic acid and n-6:n-3 ratio (P = 0·000). Owing to supplementation, milk from LPS dams had the highest DHA and ARA, but lower SCFA and medium-chain fatty acids as compared with milk from C and LP dams during early lactation, but normalised by mid-lactation. To conclude, adverse effects of restricted maternal protein intake on LC-PUFA accretion in the brain of offspring were ameliorated by alterations in maternal milk fatty acid profile due to supplementation. Results underscore the importance of LC-PUFA for protein-deficient mothers during gestation as well as lactation to achieve the optimum brain LC-PUFA status of progeny.