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Docosahexenoic acid (DHA, 22:6n-3) plays an important role in development of proper brain function in mammals. We have previously reported that DHA promotes synaptogenesis and synaptic function in hippocampal neurons while DHA-depletion in the brain due to n-3 fatty acid deficiency produces opposite effects. To gain insight into underlying molecular mechanisms, we investigated whether the brain DHA status affects the synaptic plasma membrane (SPM) proteome by using nanoLC-ESI-MS/MS and (16)O/(18)O labeling. The DHA level in mouse brains was lowered by dietary depletion of n-3 fatty acids, and SPM was prepared by differential centrifugation followed by osmotic shock. SPM proteins from DHA-adequate and depleted brains were analyzed by nanoLC-ESI-MS/MS after SDS-PAGE, in-gel digestion, and differential O(18)/O(16) labeling. This strategy allowed comparative quantitation of more than 200 distinct membrane or membrane-associated proteins from DHA-adequate or depleted brains. We found that 18 pre- and postsynaptic proteins that are relevant to synaptic physiology were significantly down-regulated in DHA-depleted mouse brains. The protein network analysis suggests involvement of CREB and caspase-3 pathways in the DHA-dependent modulation of synaptic proteome. Reduction of specific synaptic proteins due to brain DHA-depletion may be an important mechanism for the suboptimal brain function associated with n-3 fatty acid deficiency.

journal of proteome research

Effects of Docosahexaenoic Acid on Mouse Brain Synaptic Plasma Membrane Proteome Analyzed by Mass Spectrometry and <sup>16</sup>O/<sup>18</sup>O Labeling

Effects of Docosahexaenoic Acid on Mouse Brain Synaptic Plasma Membrane Proteome Analyzed by Mass Spectrometry and 16O/18O Labeling

Effects of Docosahexaenoic Acid on Mouse Brain Synaptic Plasma Membrane Proteome Analyzed by Mass Spectrometry and ¹⁶O/¹⁸O Labeling