The Yellow River, one of the very few in the Earth, originated from many dispersive palaeolakes. Taking this unique advantage, we examined the roles of palaeolake isolation vs. geological processes vs. climate in determining current fish biogeographic pattern. We reviewed available data on fish species and their geographical distribution in the river, as well as palaeolake development, geological and climatic parameters. The 138 fish species recorded in the river could be divided into 8 biogeographic regions, corresponding to the distribution of palaeolakes and respective endemic species. Through variation partitioning analysis, palaeolake isolation was the most influential factor explaining 43.6% of the total variance on the current fish distribution. The Quaternary Ice Age produced a transitional distribution for fishes from the glacier to warm water, especially for the subfamily Schizothoracinae, which showed various degrees of specialisation along altitudes. We suggested that fish biogeography in the Yellow river was basically shaped by palaeolake isolation, and further carved under serials of geologic events and contemporary climate change.