Galaxias maculatus, broadly distributed in the southern hemisphere, presents both diadromic and landlocked populations. The upstream migration of larvae and juveniles from the sea to freshwater habitats is a characteristic phenomenon of diadromous populations, but upstream migration has never been reported in those that are landlocked. The objective of this study was to establish the population dynamics of a peculiar riverine landlocked population that presents an upstream migration at the larvae–juvenile transition from the Piedra del Águila Reservoir to the Caleufú River (Argentina). A spawning season from September to November and the arrival of shoals of metamorphic larvae and juveniles from February to April to the adult habitat were coincidental with lacustrine landlocked populations, but not with diadromous populations. Growth rate and age at migration, 147 ± 22.6 days, were also similar to other landlocked and diadromous populations. The arrival of these shoals produced a 20-fold increase in fish density while the two cohorts of G. maculatus overlapped in time and space. No significant differences in morphology or vertebrae number were detected when riverine and reservoir adults were compared. The great life history plasticity of G. maculatus, shown to be even greater in our results, could be the key to explain the wide distribution of this species in the southern hemisphere. Likewise, discovering this juvenile potamodromous behaviour in a landlocked population will provide a new view for the analysis of the ways of this species’ dispersion in continental waters.