Heterogeneity in 87Sr/86Sr ratios of river-dissolved strontium (Sr) across geologically diverse environments provides a useful tool for investigating provenance, connectivity and movement patterns of various organisms and materials. Evaluation of site-specific 87Sr/86Sr temporal variability throughout study regions is a prerequisite for provenance research, but the dynamics driving temporal variability are generally system-dependent and not accurately predictable. We used the time-keeping properties of otoliths from non-migratory slimy sculpin (Cottus cognatus) to evaluate multi-scale 87Sr/86Sr temporal variability of river waters throughout the Nushagak River, a large (34,700 km2) remote watershed in Alaska, USA. Slimy sculpin otoliths incorporated site-specific temporal variation at sub-annual resolution and were able to record on the order of 0.0001 changes in the 87Sr/86Sr ratio. 87Sr/86Sr profiles of slimy sculpin collected in tributaries and main-stem channels of the upper watershed indicated that these regions were temporally stable, whereas the Lower Nushagak River exhibited some spatio-teporal variability. This study illustrates how the behavioral ecology of a non-migratory organism can be used to evaluate sub-annual 87Sr/86Sr temporal variability and has broad implications for provenance studies employing this tracer.