1. The eastern pondmussel, Sagittunio nasutus (Bivalvia: Unionida), has declined in abundance and distribution in eastern North America over the last few decades.The declines are predominantly the result of infestation by invasive dreissenid mussels and changes in habitat. The species is now considered imperilled across large portions of its distribution, especially in the Laurentian Great Lakes region.2. The genetic diversity and structure of the remnant populations in the Great Lakes region were assessed using 10 newly developed microsatellite DNA loci for S. nasutus. Understanding the remaining populations can inform future management projects and determine whether the remnant populations have experienced a genetic bottleneck or a founder effect. We hope that this will inform the conservation of other species in regions founded by, and isolated from, a more genetically diverse source population or with disjointed geographical distributions.3. Focusing on the Great Lakes region, samples (n = 428) from 62 collection sites in 28 water bodies were collected. Across the locations sampled for S. nasutus 11 genetic populations were identified, with significant genetic differentiation among them. The genetic structure of the species was assessed, with genetic similarities among populations compared and with geographical routes of colonization and gene flow evaluated. 4. Initial colonization from the Atlantic coast into Lake Erie and Lake Ontario is evident, followed by colonization events into nearby inland rivers and lakes. Analyses found evidence of inbreeding in all but one population and evidence of past genetic bottlenecks or strong founder effects in all but four populations. This study deepens our understanding of the genetic past and present of this imperilled species, providing conservation suggestions for the future management of the species. K E Y W O R D S founder effects, genetic bottlenecks, Ligumia nasuta, management units, microsatellite DNA, phylogeography, population structure, rescue effect, Sagittunio nasutus, Unionidae