Immunohistochemical localization of V-ATPases in rat spermatids

The sperm acrosome exhibits a low pH. However, the mechanism of acidification in the acrosome remains unclear. Vacuolar-type proton ATPase (V-ATPase) has been shown to play a principle role in generating and maintaining the acidity of organelles such as lysosomes and endosomes. In this study, we examined whether V-ATPase is localized in the acrosome membranes using immunohistochemical techniques. Sections of rat testis were immunostained using antibodies against V-ATPase. Under light microscopic observation, the perinuclear region in spermatids at an early stage of development was heavily immunostained. At the electron microscopic level, gold particles showing the presence of V-ATPase were localized to the acrosome membranes in the developing spermatids. V-ATPase was also localized to the membrane of vesicles locating between the trans-Golgi area and the acrosome. These observations suggest that V-ATPase may play a role in acrosome acidification.