The Nashville crayfish is endemic to the Mill Creek watershed in Davidson and Williamson counties, Tennessee. The species was thought to occur historically in a few locations outside of the Mill Creek watershed, including Big Creek in Giles County (Elk River drainage), the South Harpeth River in Davidson County (Harpeth River drainage), and Richland Creek in Davidson County (Cumberland River drainage) (USFWS 1987). The Big Creek and South Harpeth River records are believed to be the result of “bait bucket” introductions. The species was thought to be native to Richland Creek and displaced by a more competitive crayfish species, the bigclaw crayfish (O. placidus). However, specimens of Nashville crayfish (O. shoupi) collected from Richland Creek were misidentified and the collections were annotated as the bigclaw crayfish (USWFS 1989).

Biologists conducting the pre-listing status survey for the species surveyed 148 streams in central Tennessee (Korgi and O’Bara 1985). Streams surveyed were located in the Collins River drainage, Stones River drainage, Caney Fork River drainage, Cumberland River drainage, Red River drainage, Mill Creek drainage, Harpeth River drainage, and Elk River drainage. Nashville crayfish were only found in Mill Creek and its tributaries.