Dams fragment watersheds and prevent anadromous fishes from reaching historic spawning habitat. Sedgeunkedunk Stream, a small tributary to the Penobscot River (Maine), has been the focus of efforts to reestablish marine-freshwater connectivity and restore anadromous fishes via the removal of two barriers to fish migration. Currently, Petromyzon marinus (Sea Lamprey) is the only anadromous fish known to spawn successfully in the stream downstream of the lowermost dam. Here, we describe the distribution and abundance of a spawning population of Sea Lamprey in Sedgeunkedunk Stream, prior to and in anticipation of habitat increase after the completion of one barrier removal. In 2008, we estimated the abundance of Sea Lamprey and its nests using daily stream surveys and an open-population mark-recapture model. We captured 47 Sea Lamprey and implanted each with a PIT tag so that we could track movements and nest associations of individual fish. The spawning migration began on 18 June, and the last living individual was observed on 27 June. We located 31 nests, distributed from head-of-tide to the lowermost dam; no spawners or nests were observed in the tidally influenced zone or upstream of this dam. Mean longevity in the stream and the number of nests attended were correlated with arrival date; early migrants were alive longer and attended more nests than later migrants. Males were more likely to be observed away from a nest, or attending three or more nests, than were females, which attended usually one or two nests. We observed a negative association between nest abundance and substrate cover by fine sediment. Based on their observed movements in the system, and the extent of their habitat use, we anticipate that spawning Sea Lamprey will recolonize formerly inaccessible habitat after dam removals.