Diet and sexual maturation were examined in the winter–spring cohort of the neon flying squid, Ommastrephes bartramii, during its southward migration through the Kuroshio–Oyashio transition region of the western North Pacific. The main prey items are micronektonic animals and small pelagic fish, which were abundantly distributed throughout the study area. Among the prey species, O. bartramii was dependent on the Japanese anchovy Engraulis japonicus, which also migrate from the northern to southern transition region during the winter, and the micronektonic squid Watasenia scintillans throughout the study period. Other common prey items were Gonatopsis borealis, Diaphus theta, and Ceratoscopelus warmingii from October to November or December, and Tarletonbeania taylori after December. These dietary changes can be explained by the difference in the seasonal north–south migration patterns of the predator and prey species. Male sexual maturation progressed throughout the season, and most individuals were fully mature in January. In contrast, most of the females were immature throughout the study period. The feeding strategy of the squid in relation to their seasonal north to south migration and sexual maturation was discussed.