The Histochemical Demonstration of Carbonic:Anhydrase in the Osteoclast

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Three histochemical methods for demonstration of CAH, i. g., Häusler's, Kurata's, and Coquoin-Carnot's methods are compared in a model experiment, using a crude preparation of CAH. Häusler's method gives the most constant and clearcut result, but two other methods are not good. For the fixatives, chilled actone, ethanol and methanol are all excellent, but neutral formlin is unsatisfactory.<BR>When Diamox 100mg% is added to the incubating medium, or the sample is pretreated at 90°C for 2 minutes, this reaction is completely inhibited, suggesting this reaction is a specific one for CAH.<BR>Of various decalcifying fluids, whose effect on this reaction is tested in the model experiment, Lorch's citrate-HCl buffer after acetone fixation is the most suitable.<BR>In undecalcified bone sections, osteoclasts are positive in contrast with the negative reaction of osteoblasts. This positive reaction of osteoclasts, which is completely inhibited by Diamox, is to be ascribed not to the bone salt phagoytized in them, but to CAH.<BR>Coexistence of CAH with Ac. P-ase in the osteoclast may be considered to indicate this enzyme's intracellular pH regulative function.
日本組織細胞化学会の論文