Accurate distribution data are critical to the development of conservation and management strategies for imperiled species, particularly for narrow endemics with life history traits that make them vulnerable to extinction. Medionidus walkeri is a rare freshwater mussel endemic to the Suwannee River Basin in southeastern North America. This species was rediscovered in 2012 after a 16-year hiatus between collections and is currently proposed for listing under the Endangered Species Act. Our study fills knowledge gaps regarding changes in distribution and early life history requirements of M. walkeri. Spatiotemporal changes in M. walkeri distribution were displayed using a conservation status assessment map incorporating metadata from 98 historical (1916–1999) and 401 recent (2000–2015) site surveys from museums and field notes representing records for 312 specimens. Recent surveys detected M. walkeri only in the middle Suwannee subbasin (n = 86, 22 locations) and lower Santa Fe subbasin (n = 2, 2 locations), and it appears the species may be extirpated from 67% of historically occupied 10-digit HUCs. In our laboratory experiments, M. walkeri successfully metamorphosed on Percina nigrofasciata (56.2% ± 8.9) and Etheostoma edwini (16.1% ± 7.9) but not on Trinectes maculatus, Lepomis marginatus, Notropis texanus, Noturus leptacanthus, Etheostoma fusiforme, or Gambusia holbrooki. We characterize M. walkeri as a lure-displaying host fish specialist and a long-term brooder (bradytictic), gravid from fall to early summer of the following year. The early life history and distribution data presented here provide the baseline framework for listing decisions and future efforts to conserve and recover the species.