The brown trout populations of the Baltic Sea region have been drastically affected by various human activities during the past century. Due to their propensity to home to their natal site to spawn and their tendency to evolve local adaptations, populations may be genetically differentiated in water systems where no physical barriers preventing interbreeding exist. Consequently, identification of management units, a prerequisite for appropriate conservation and management planning, cannot necessarily be deduced from the physical properties of the habitat. In this study, microsatellite markers were employed to assess the spatio-temporal genetic structuring of inter-connected brown trout populations from a river-system in Northwest Russia. Populations were found to be genetically differentiated from each other (global F ST 0.06) and the genetic structuring within the river to follow an isolation by distance pattern. Indications of temporal stability were found in some populations, however others appeared to be temporally unstable suggesting differences in the demographic forces affecting the populations. Based on the observed isolation by distance pattern of genetic differentiation, preserving several breeding sites spaced evenly throughout the river-system would appear to be more appropriate than focussing conservation effort on any single stretch of the river.