In-stream barriers pose threats to fishes, including habitat loss, constraints on migration, and reduced connectivity between populations. Despite many negative consequences, barriers can serve to protect native species by limiting the spread of invasive species. For example, in the Laurentian Great Lakes, physical barriers have long been used to control invasive sea lamprey (Petromyzon marinus) populations by limiting access to potential upstream spawning and rearing habitat. Selective fish passage systems could solve this management trade-off, termed the “connectivity conundrum”, but must efficiently pass multiple native or desirable species while blocking invasive species. Designing such fish passage systems requires an understanding of the attribute dimensions of the fish community, specifically, the phenology, morphology, physiology, and behaviour of each species. Here, we describe the first comprehensive collection of sortable attributes associated with fish passage. The integrated database consists of 21 biological attributes that influence the movement and passage of 220 species in the Great Lakes, including native species, established non-native species, and unestablished but potentially invasive fishes. Data coverage varies with species, taxonomic orders, and attribute dimensions. Behavioural attributes were typically underrepresented in the literature, and the ecology of potential invaders was not well understood. The synthesis described herein is a critical step towards a holistic approach to fish passage design and may help to inform management actions related to population connectivity. The database is openly accessible online and is expected to be updated periodically.