Mountain Whitefish Prosopium williamsoni were historically common throughout much of the U.S. Intermountain West. However, within the last decade Mountain Whitefish have exhibited population-level declines in some rivers. In the Madison River, Montana, anecdotal evidence indicates Mountain Whitefish abundance has declined and the population is skewed toward larger individuals, which is typically symptomatic of recruitment problems. Describing reproductive development, spawning behavior, and juvenile distribution will form a foundation for investigating mechanisms influencing recruitment. We collected otoliths and gonadal samples from fish of all size-classes to characterize fecundity, age at maturity, and spawning periodicity. We implanted radio tags in mature Mountain Whitefish and relocated tagged fish in autumn 2012–2014. Timing of spawning was determined from spawning status of captured females and from density of eggs collected on egg mats. In spring 2014, we seined backwater and channel sites to describe age-0 whitefish distribution. Mountain Whitefish were highly fecund (18,454 eggs/kg body weight) annual spawners, and age at 50% maturity was 2.0 years for males and 2.6 years for females. In 2013 and 2014, spawning occurred between the third week of October and first week of November. During spawning, spawning adults and collected embryos were concentrated in the downstream 26 km of the study site, a reach characterized by a complex, braided channel. This reach had the highest CPUE of age-0 Mountain Whitefish, and the percentage of spawning adults in the 25 km upstream from a sampling site was positively associated with juvenile CPUE. Within this reach, age-0 Mountain Whitefish were associated with silt-laden backwater and eddy habitats. Future investigations on mechanisms influencing recruitment should be focused on the embryological phase and age-0 fish.