The Paddlefish (Polyodon spathula), a large-bodied and highly migratory species of large river systems, has experienced population declines or extirpation in parts of its native range. As an effort to reestablish a Paddlefish population in the Ohio River of West Virginia, the West Virginia Division of Natural Resources has stocked fingerling Paddlefish in the previous two decades. Post-stocking studies could assess the status of the reintroduced population, which includes collecting information on their seasonal movements and dam passage frequency through navigation dams of the Ohio and Kanawha Rivers. Using acoustic telemetry techniques, we monitored seasonal movements and assessed dam passage of five Paddlefish in the Robert C. Byrd Pool of the Ohio and Kanawha rivers. Paddlefish were highly mobile with two individuals using the entire R.C. Byrd Pool during the 11-month study period. Most movements occurred during the expected spawning period of April and May, where all individuals frequented the upper section of the pool in the Kanawha River. Downstream passage was documented for one individual at the R.C. Byrd lock during July 2023, but no evidence of upstream passage was documented for the locks at Racine (Ohio River) or Winfield (Kanawha River). Tagging of additional Paddlefish would be beneficial to increase sample size for future assessments and continue efforts toward the long-term conservation and management of Ohio River Paddlefish.