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Accumulation of docosahexaenoic acid (DHA; 22:6n-3) in brain and eyes during development has been demonstrated in fish but it is not clear whether liver or neural tissues themselves are of greater importance in the biosynthesis of DHA from dietary 18:3n-3. In the present study, we investigated the in vivo metabolism of intraperitoneally injected [1-14C]18:3n-3 in liver, brains and eyes of young juvenile fish. Metabolism was followed over a 48h time-course in order to obtain dynamic information that could aid the elucidation of the roles of the different tissues in the biosynthesis and provision of DHA from dietary 18:3n-3. The study was performed in both a freshwater fish, rainbow trout Oncorhynchus mykiss L and a marine fish, gilthead sea bream Sparus aurata L to determine the effect that low or limitingΔ5-desaturase activity may have in this process. As expected, the results showed that although the sea bream incorporated more 18:3n-3 into its lipids, metabolism of the incorporated fatty acid by de saturation and elongation was generally greater in the trout. In liver, the percentages of radioactivity recovered in tetraene and pentaene products were greater in trout than in sea bream although there was no difference in hexaenes. In contrast, the re covery of radioactivity in DHA was significantly greater in brain in trout compared to sea bream. In both species, the percentage of radioactivity recovered in desaturated/elongated products was much lower in liver than in brains and eyes, but that percentage increased over the 48h time-course. In trout though, the highest percentages of desaturated products in brain and eye were observed after 12 and 24h, respectively. However in sea bream the highest percentages of desaturated products in the neural tissues were observed after 24-48h. Radioactivity was recovered in 24:5n-3 and 24:6n-3, intermediates in the Δ4-independent ("Sprecher shunt") pathway for the synthesis of DHA, in both species, especially in the brain and eyes. Overall, although the results cannot eliminate a role for liver in the biosynthesis and provision of DHA for developing neural tissues in fish, they suggest that DHA can be synthesised in fish brain and eye in vivo.