Stimulation of carbonic anhydrase in osteoclasts by parathyroid hormone

Changes in the acidity of osteoclasts were evaluated by direct measurement of the fluorescent intensity of osteoclasts exposed to acridine orange, a fluorescent weak base which becomes concentrated in acid-containing subcellular compartments. Parathyroid hormone (PTH) produced dose-dependent increases in fluorescent intensity; maximal increases in intensity occurred at doses between 3 and 10 micrograms PTH/ml of culture medium. Acetazolamide, a potent inhibitor of carbonic anhydrase, inhibited the increase in fluorescence induced by PTH, but this drug was less effective in reducing fluorescence in maximally than in submaximally stimulated osteoclasts, indicating that either more enzyme or more resistant enzyme was present in the PTH-stimulated cells. Because increased fluorescence of acridine orange is a sign of greater acidity, these results suggest that (1) PTH stimulates the acidity of osteoclasts, (2) carbonic anhydrase activity is necessary for maximum acidity, and (3) carbonic anhydrase is activated by PTH.