AbstractReproductive isolation is pivotal to maintain species separation and it can be achieved through a plethora of mechanisms. In addition, the development of barriers to gamete interaction may drive speciation. Such barriers to interspecific gamete interaction can be prezygotic or postzygotic. Considering the great diversity in animal species, it is easy to assume that regulation of the early steps of fertilization is critical to maintain species identity. One prezygotic mechanism that is often mentioned in the literature is that gamete interaction is limited to gametes of the same species. But do gametes of all animals interact in a species‐specific way? Are gamete interactions completely species‐specific or perhaps just species‐restricted? In species in which species‐restrictions have been described, is the interspecies barrier at one major step in the fertilization process or is it a combination of partially restricted steps that together lead to a block in interspecific fertilization? Are the mechanisms used to avoid interspecific crosses different between free‐spawning organisms and those with internal fertilization? This review will address these questions, focusing on prezygotic barriers, and will describe what is known about the molecular biology that may account for species‐limited gamete recognition and fertilization. Mol. Reprod. Dev. 73: 1422–1429, 2006. © 2006 Wiley‐Liss, Inc.