The ability to breed zebrafish in the laboratory goes a long way in determining the success or failure a research program. The complexity of zebrafish reproductive behavior and spawning is often vastly underestimated. This presentation will provide attendees with an in-depth look at current understanding of zebrafish reproduction, as well as a comprehensive overview of some of the most critical factors impacting the breeding performance of this important laboratory animal. This information will help technicians, managers, researchers, and veterinarians to both troubleshoot existing breeding problems and build successful zebrafish breeding programs at their own institutions over the long-term.