A substantial body of literature exists about depth distribution of fish in oceans, lakes and reservoirs, but less is known about fish depth distribution in large rivers. Most of the emphasis on fish distributions in rivers has focused on longitudinal and latitudinal spatial distributions. Knowledge on depth distribution is necessary to understand species and community habitat needs. Considering this void, our goal was to identify patterns in fish benthic distribution along depth gradients in the Lower Mississippi River. Fish were collected over 14 years in depths down to 27 m. Fish exhibited non-random depth distributions that varied seasonally and according to species. Species richness was highest in shallow water, with about 50% of the 62 species detected no longer collected in water deeper than 8 m and about 75% no longer collected in water deeper than 12 m. Although richness was highest in shallow water, most species were not restricted to shallow water. Rather, most species used a wide range of depths. A weak depth zonation occurred, not as strong as that reported for deep oceans and lakes. Larger fish tended to occur in deeper water during the high-water period of an annual cycle, but no correlation was evident during the low-water period. The advent of landscape ecology has guided river research to search for spatial patterns along the length of the river and associated floodplains. Our results suggest that fish assemblages in large rivers are also structured vertically.