Sperm morphology can be highly variable among individuals and across species, but less is known about its variation among populations. Within the past 20–80 yr, several species of Anolis lizards have been introduced to Miami, Florida, USA from different source islands in the Caribbean, thereby permitting comparisons of sperm morphology between native and introduced populations of multiple species. We collected sperm samples from native populations of Anolis sagrei (Bahamas), Anolis distichus (Dominican Republic), and Anolis cristatellus (Puerto Rico) and compared them to samples from introduced populations of each species that are now sympatric in Miami. In each of these three species, lizards from introduced populations had sperm with shorter tails and larger midpieces relative to lizards from native populations. We also measured testis size in A. distichus and A. cristatellus and found that introduced populations of each species had smaller testes for a given body size relative to their native counterparts. The consistency of these differences across species argues against random genetic drift as an explanation, suggesting instead that sperm morphology and testis size may exhibit predictable phenotypic plasticity or genetic adaptation in response to the process of introduction and/or the shared local environment in Florida. Though these population differences in male reproductive physiology and morphology may be repeatable, their underlying causes require further study.