1. Defining functional connectivity between habitats in spatially heterogeneous landscapes is a particular challenge for small-bodied aquatic species. Traditional approaches (e.g. mark–recapture studies) preclude an assessment of animal movement over the life cycle (birth to reproduction), and movement of individuals may not represent the degree of gene movement for fecund species. We investigated the degree of habitat connectivity (defined as the exchange of individuals and genes between mainstem and tributary habitats) in a stream brook trout (Salvelinus fontinalis) population using mark–recapture [passive integrated transponder (PIT) tags], stationary PIT-tag antennae and genetic pedigree data collected over 4 years (3425 marked individuals). We hypothesised that: (i) a combination of these data would reveal higher estimates of animal movement over the life cycle (within a generation), relative to more temporally confined approaches, and (ii) movement estimates of individuals within a generation would differ from between-generation movement of genes because of spatial variation in reproductive success associated with high fecundity of this species. Over half of PIT-tagged fish (juveniles and adults) were recaptured within 20 m during periodic sampling, indicating restricted movement. However, continuous monitoring with stationary PIT-tag antennae revealed distinct peaks in trout movements in June and October–November, and sibship data inferred post-emergence movements of young-of-year trout that were too small to be tagged physically. A combination of these methods showed that a moderate portion of individuals (28–33%) moved between mainstem and tributary habitats over their life cycle. Patterns of reproductive success varied spatially and temporally. The importance of tributaries as spawning habitat was discovered by accounting for reproductive history. When individuals born in the mainstem reproduced successfully, over 50% of their surviving offspring were inferred to have been born in tributaries. This high rate of gene movement to tributaries was cryptic, and it would have been missed by estimates based only on movement of individuals. This study highlighted the importance of characterising animal movement over the life cycle for inferring habitat connectivity accurately. Such movements of individuals can contribute to substantial gene movements in a fecund species characterised by high variation in reproductive success.