Connectivity is a foundational concept in ecology and conservation and was the organising theme for the 2022 Annual Meeting of the Southeastern Fishes Council, a professional organisation dedicated to the study and conservation of freshwater fishes native to the southeast region of the United States (US). We introduce a Special Contribution of six papers selected from presentations at that meeting that illustrate perspectives on connections created by fish migration and dispersal, evolved life histories and habitat affinities and interspecific facilitation. Although focused on streams of the southeast US, each of these topics is broadly relevant to freshwater fish conservation, particularly with respect to causes and consequences of migratory fish depletion, population fragmentation and species declines. Many other connections relevant to the ecology and conservation of freshwater fishes remain relatively unexplored but could substantively advance conservation. We highlight the potential that species evolutionary histories, that is connections through time, reconstructed using species distributions and phylogenies may improve predictions of species responses to environmental change. Identifying species interdependencies, including undiscovered interactions that support survival or reproduction, could provide insights into how species losses may cascade as aquatic communities unravel. Finally, efforts to elucidate diverse connections between people and freshwater biodiversity, particularly where fisheries are historic and streams mostly go unnoticed, may prove essential to building public support for conservation measures. A research agenda anchored on ‘biodiversity connections’ has the potential to advance ecological understanding and public engagement, elements essential to conserving freshwater fishes.