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Reproductive characteristics are one of the key factors for understanding population dynamics in fishes. This study describes the spawning habitat of Pacific bluefin tuna (Thunnus orientalis) in the Kuroshio and the Kuroshio–Oyashio transition area. A total of 551 adult fish (fork length > 120 cm) were collected during 1994–2007 (except in 2001 and 2003) and 2016. Based on a histological analysis of the gonads, we concluded that actively spawning females were mainly present in late May to early July. Relative batch fecundity was significantly lower than that reported in the Sea of Japan but was similar to that observed in the Nansei area. Whereas most spawning adults in the Sea of Japan are 3–6 years old and those in the Nansei area are over 10 years old, the majority of the specimens in this study ranged from 160 to 180 cm in length, which corresponds to 6–8 years of age. Our results indicate that Pacific bluefin tuna spawn in the study area and provide important data for understanding the spatiotemporal differences in spawning strategy with growth in this species.