Lampreys typically spawn in riffle habitats during the spring. Spawning activity and diel (i.e., during daylight and at night) behavioral patterns are initiated when spring water temperatures increase to levels that coincide with optimal embryologic development. Nests are constructed in gravel substrate using the oral disc to move stones and the tail to fan sediment out of the nest. Spawning habitat used by individual species is generally a function of adult size, where small-bodied species construct nests in shallower water with slower flow and smaller gravel than large-bodied species. The mating system of lampreys is primarily polygynandrous (i.e., where multiple males mate with multiple females). Lamprey species with adult total length less than 30 cm generally spawn communally, where a nest may contain 20 or more individuals of both sexes. Lamprey species with adult sizes greater than 35 cm generally spawn in groups of two to four. Operational sex ratios of lampreys are highly variable across species, populations, and time, but are generally male biased. The act of spawning typically starts with the male attaching with his oral disc to the back of the female’s head; the male and female then entwine and simultaneously release gametes. However, alternative mating behaviors (e.g., release of gametes without paired courtship and sneaker males) have been observed. Future research should determine how multiple modalities of communication among lampreys (including mating pheromones) are integrated to inform species recognition and mate choice. Such research could inform both sea lamprey control strategies and provide insight into possible evolution of reproductive isolation mechanisms between paired lamprey species in sympatry.