Responses of benthic macroinvertebrates along gradients of urban intensity were investigated in nine metropolitan areas across the United States. Invertebrate assemblages in metropolitan areas where forests or shrublands were being converted to urban land were strongly related to urban intensity. In metropolitan areas where agriculture and grazing lands were being converted to urban land, invertebrate assemblages showed much weaker or nonsignificant relations with urban intensity because sites with low urban intensity were already degraded by agriculture. Ordination scores, the number of EPT taxa, and the mean pollution‐tolerance value of organisms at a site were the best indicators of changes in assemblage condition. Diversity indices, functional groups, behavior, and dominance metrics were not good indicators of urbanization. Richness metrics were better indicators of urban effects than were abundance metrics, and qualitative samples collected from multiple habitats gave similar results to those of single habitat quantitative samples (riffles or woody snags) in all metropolitan areas. Changes in urban intensity were strongly correlated with a set of landscape variables that was consistent across all metropolitan areas. In contrast, the instream environmental variables that were strongly correlated with urbanization and invertebrate responses varied among metropolitan areas. The natural environmental setting determined the biological, chemical, and physical instream conditions upon which urbanization acts and dictated the differences in responses to urbanization among metropolitan areas. Threshold analysis showed little evidence for an initial period of resistance to urbanization. Instead, assemblages were degraded at very low levels of urbanization, and response rates were either similar across the gradient or higher at low levels of urbanization. Levels of impervious cover that have been suggested as protective of streams (5–10%) were associated with significant assemblage degradation and were not protective.