We assessed temporal variation in mercury (Hg) concentrations of threespine stickleback (Gasterosteus aculeatus) from Agattu Island, Aleutian Archipelago, Alaska. Total Hg concentrations in whole-bodied stickleback were measured at two-week intervals from two sites in each of two lakes from June 1 to August 10, 2011 during the time period when lakes were ice-free. Across all sites and sampling events, stickleback Hg concentrations ranged from 0.37–1.07 µg/g dry weight (dw), with a mean (± SE) of 0.55±0.01 µg/g dw. Mean fish Hg concentrations declined by 9% during the study period, from 0.57±0.01 µg/g dw in early June to 0.52±0.01 µg/g dw in mid-August. Mean fish Hg concentrations were 6% higher in Loon Lake (0.56±0.01 µg/g dw) than in Lake 696 (0.53±0.01 µg/g dw), and 4% higher in males (0.56±0.01 µg/g dw) than in females (0.54±0.01 µg/g dw). Loon Lake was distinguished from Lake 696 by the presence of piscivorous waterbirds during the breeding season. Mercury concentrations in stickleback from Agattu Island were higher than would be expected for an area without known point sources of Hg pollution, and high enough to be of concern to the health of piscivorous wildlife.