Sufficient oxygen and nutrient supplies to the retina are dependent on a correct development of blood vessels. Polyunsaturated fatty acids (PUFAs) were shown to modulate factors involved in tissue vascularization. A congenital deficiency in ether‐linked phospholipids also termed as “plasmalogens” was shown to lead to abnormal ocular vascularization. Because plasmalogens are considered as reservoirs of PUFAs, we investigated whether they may influence retinal vascular development through the release of fatty acids by a specific phospholipase A2 (PLA2). Studies on mouse models of plasmalogen deficiency and of retinal PLA2 inhibition have shown that plasmalogens regulate retinal angiogenesis during early phases of retinal vascular development by influencing glial activity and primary astrocyte template formation. Subsequent modifications in the expression of genes involved in retinal angiogenesis impacted endothelial cell proliferation and retinal vessel outgrowth. These early defects in retinal vascular development led to a disorganized and dysfunctional retinal vascular network at adult age. These data suggest a crucial role of plasmalogens in retinal vascularization processes through a signalization involving PUFAs and PLA2.