The Macquarie perch (Macquaria australasica) is a threatened fish species that inhabits rivers and impoundments in south-eastern Australia. Previous studies have shown that Macquarie perch in impoundments exhibit synchronised upstream spawning migrations to shallow, fast-flowing habitats in the lower reaches of inflowing streams. There has been little study of movement behaviours of entirely riverine populations of Macquarie perch despite this being the species’ natural habitat. Here, radio-telemetry is used to test the hypothesis that riverine populations exhibit synchronised migrations during the spawning season. Thirty Macquarie perch in the Yarra River, Victoria, a translocated population outside of the species’ natural range, were radio-tagged before the late spring–early summer spawning season and their movements followed over a 10-month period (May 2011 to February 2012). Tagged fish typically occupied restricted reaches of stream (<450 m). Sixteen of the fish undertook occasional upstream or downstream movements (∼250–1000 m) away from their usual locations, particularly associated with large flow variations during the spawning season. There was no evidence of synchronised migratory behaviour or movement of multiple fish to specific locations or habitats during the spawning season. Whilst further research over more years is needed to comprehensively document the spawning-related behaviours of riverine Macquarie perch, our study demonstrates that management of riverine populations of this threatened species cannot necessarily be based on the model of spawning behaviour developed for lacustrine populations.