The histochemical localization of ATPase, cholinesterase and acid phosphatase activity in (Diptera, Culicidae) larvae using a methacrylate embedding technique

Abstract

A method is described for the demonstration of ATPase, Cholinesterase and acid phosphatase activity in thin sections of mosquito larvae fixed in 1:9 v/v mixture of acetone and 10% neutral buffered formalin and embedded in hydroxyethyl methacrylate (HEMA). ATPase activity, observed as a black brown precipitate, was found in the brush border of gastric caeca and microvilli of columnar epithelial cells of the hind gut and Malpighian tubules. Some basal cell membrane activity could also be seen. Cholinesterase activity was found in thoracic and abdominal ganglia. The reaction product had a fine particulate appearance and predominated in the axonal processes. Azo dye reaction product indicative of acid phosphatase activity was found in the epithelial cells of the midgut and gastric caeca. Lysosomal and extra-lysosomal activity was observed, the larger secondary lysosomal sources predominating in the perinuclear region. The fixation regime and embedding procedure outlined has enabled a sub-cellular localization of enzymatic activities which is superior to that obtainable with conventional procedures. © 1990.