The presence and localization of acrosin and hyaluronidase have been detected by immunocytochemistry in animals belonging to different phyla and characterized by the different structure of the acrosomal complex. Acrosin is widely distributed from Insecta to Echinodermata and Vertebrata; hyaluronidase has a similar diffusion, but seems to be absent from an Insect and from Echinodermata. The presence of extraacrosomal layer and perforatorium seems not to be related to that of the two enzymes, that are absent from Teleost species devoid of acrosome or having only a pseudoacrosome.