Published species lists were analysed to determine the contributions of dispersal, habitat preference, river channel size, body size, and glacial history to large-scale patterns in freshwater fish species richness in North America, north of central Mexico. Total species richness declines to the north and west but the pattern for endemics differs from that of widespread species. Mississippi Basin regions are more species rich than more isolated, coastal, regions. Richness declines more rapidly with increasing latitude in riverine specialist than in habitat generalist species. Levels of endemism are greatest in species found in small- to medium-sized river channels. The strong Rapoport effect, more marked in migratory than resident species, is correlated with habitat preference, channel size, and glacial history. Body size increases with latitude, largely as a result of a trend from small resident to large migrant species. In unglaciated regions, ancestral species survived in large habitats because these are longer-lived, more extensive, less isolated and more stable than headwaters, permitting larger populations and lower extinction levels. Reduced levels of gene flow in small, peripheral, channels isolated by larger downstream habitats have resulted in the production of many, small range, small-bodied species. The latitudinal richness gradient is a consequence of speciation and extinction events in unglaciated faunas and an increasing domination of faunas by generalist, large bodied, large channel, recolonizing species in more northern regions. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100, 46–61.