Membrane Molecular Species Remodeling as a Signature of ω-3 Fatty Acid Action in Cultured Neural Cells

Omega-3 polyunsaturated fatty acids (ω3 PUFAs) are critical structural components of neuronal membranes, yet the molecular specificity of their incorporation within neural cells remains incompletely defined. We integrated untargeted and targeted lipidomics with lipid ontology analysis and coarse-grained membrane simulations to characterize remodeling in primary rat cortical neurons and neuron-astrocyte co-cultures following supplementation with docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or docosapentaenoic acid (DPA). Each ω3 PUFA produced a distinct lipidomic signature. DHA showed the most consistent incorporation, selectively enriching phosphatidylethanolamine (PE) species-particularly PE(18:0/22:6) and PE(18:1/22:6)-associated with membrane curvature and organelle organization. Ontology analysis linked DHA supplementation to intrinsic curvature-related membrane features, and membrane simulations demonstrated enhanced collective bilayer bending without substantial changes in overall membrane thickness. EPA preferentially increased EPA-containing PE species without elevating DHA levels, whereas DPA effects were variable and culture-dependent, indicating selective metabolic handling of individual ω3 species. Differences between neurons and neuron-astrocyte co-cultures underscore the importance of cellular context in ω3-driven remodeling. By resolving ω3 incorporation at molecular species resolution and linking compositional changes to predicted membrane behavior, this study provides a structural framework for understanding how dietary ω3 fatty acids may influence neuronal membrane organization and cellular resilience.