The importance of fish spawning habitat in channels connecting the Great Lakes to fishery productivity in those lakes is poorly understood and has not been adequately documented. The Detroit River is a reputed spawning and nursery area for many fish, including walleye (Sander vitreus) that migrate between adjacent Lakes Erie and St. Clair. During April–May 2004, near the head of the Detroit River, we collected 136 fish eggs from the bottom of the river on egg mats. We incubated the eggs at the Great Lakes Science Center until they hatched. All eleven larvae that hatched from the eggs were identified as walleye. These eggs and larvae are the first credible scientific evidence that walleye spawn in the Detroit River. Their origin might be a stock of river-spawning walleye. Such a stock of walleye could potentially add resilience to production by walleye stocks that spawn and are harvested in adjacent waters.