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Summary.: Ram spermatozoa were washed with hypotonic tris-HCl buffer and obtained almost free of seminal plasma and cytoplasmic droplets. Acrosomes were dislodged by incubating the spermatozoa with a cationic detergent (Hyamine 2389). The acrosomal preparation and the buffer washings were examined for the following lysosomal enzymes: acid phosphatase, aryl sulphatase, β-N-acetylglucosaminidase, phospholipase A and proteases. All enzyme activities were detected, both in washings and in acrosomal preparations. The levels of activity in the latter were much higher than could be expected on the assumption that all activities were due to contamination by washings. Protease activity was greatest at pH 7·5.
After vital staining with Euchrysine 3R, acrosomal fluorescence in ram, bull, boar and human spermatozoa is not very conspicuous. However, acrosomes of guinea-pig, hamster and several rodents show the brilliant orange-red fluorescence typical of lysosomes. As spermatids mature, red-fluorescing granules around the Golgi zone condense to form the red-fluorescent pro-acrosomes.
Acid phosphatase was detectable histochemically in granules, pro-acrosomes, acrosomes and the pellets obtained by high-speed centrifugation of acrosomal preparations. At all developmental stages, histochemical tests for bromochloroindoxyl acetate esterase showed that this enzyme was present only in the acrosome.
The evidence suggests that the acrosome is a specialized lysosome which evolved to facilitate fertilization in multicellular organisms.
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