Dozens of well-preserved fossil burrow systems in upper Miocene sedimentary rocks of the Ogallala Group at a site in east-central Nebraska record rodent behavior and the subsurface ecology of grasslands just as the modern Great Plains was developing. These burrow systems include one to four entrance or exit tunnels, large underground nesting chambers at depths of several decimeters below ancient land surfaces, and incisor grooves on the walls. Tunnels average 89.8 mm in diameter, a value similar to the burrow diameters of multiple living North American rodents. Chambers vary in shape and typically exceed 500 mm in length; some attain 1000 mm in length. Living marmotine ground squirrels (tribe Marmotini) construct burrow systems of varying degrees of complexity, but they do not engage in shallow subsurface foraging. Extinct members of this group were the most likely excavators of the fossil burrows. In contrast, extant pocket gophers (family Geomyidae) and, presumably their fossil relatives, are obligate subterranean animals that produce linked deep and shallow burrow subsystems, the latter representing their chief foraging strategy. Our results raise issues regarding the relationships between the architecture of fossil rodent burrow systems and aspects of rodent behavior and life history, such as litter size, developmental rates, seasonal torpor, hibernation, and sociality in grasslands. An improved understanding of the burrowing behaviors of ancient rodents will highly complement the growing body of knowledge about the development of grasslands on Earth over time, but truly ichnological analyses of the burrows and burrowing behaviors of extant rodents are much needed.