Purpose: Omega-3 polyunsaturated fatty acids (ω3) are key components in nervous structures but their dietary intakes in the overall population are often below nutritional requirements. A chronic deficiency in ω3 is recognized to be associated with functional impairment of the retina. At the opposite, ω3 supplementation is associated with a reduced risk for AMD. The consequences of ω3 deficiency on other eye structures than the retina, such as ciliary bodies, are scarce. The purpose of our study was to compare the response of the retina and ciliary bodies to dietary ω3 deficiency in terms of fatty acid profile and eye functionality. Methods: Two successive generations of Lewis rats (G1 and G2) were obtained under either a standard or ω3-deficient diet. Intraocular pressure (IOP) was measured by rebound tonometry throughout the experiment. Retinal functionality was assessed by scotopic electroretinography (ERG). Gas chromatography was used to determine the fatty acid profile of the ciliary bodies and retina. Results: A 2-fold fall in DHA content of the retina was observed in ω3-deficient G1 animals. This decrease was accentuated in G2 (-66%) and counterbalanced by an increase in DPAω6 in the retina. The b-wave amplitude of the ERG was decreased by 50% at 9mcds/m² in ω3-deficient rats. In ciliary body DHA was reduced by 80% in ω3-deficient in G1 animals but not in G2. Meanwhile, animals from the ω3-deficient diet had increased IOP (18 vs 12mmHg, p<0.0001). Conclusion: The crucial role of ω3 in retinal function was confirmed. The most relevant finding from our study is the rise in IOP, the major risk factor for glaucoma, which was observed in animals reared under dietary deficiency in ω3.