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Cyclic nucleotides (cAMP and cGMP) and prostaglandin E (PGE) have been implicated as possible mediators of the effects of external stimuli on bone cells. The objective of this study was to determine changes in relative levels of these substances in mineralized tissue cells in response to mechanical and electrical stimuli, by the use of a combined immunohistochemical-microphotometric procedure. Canine teeth of eight 10-12 month-old female cats were tipped distally with 80 g force for either 1 h or 14 days. After 1 h, a slight elevation of staining intensity in alveolar bone osteoblasts and periodontal ligament (PDL) cells was observed at sites of tension and compression. After 14 days of treatment, this effect was markedly increased. Fifteen female cats, 10-12 months old, received electric stimulation (20 micron amperes d.c.) to the gingiva of 1 maxillary canine for 1, 5, 15, 30, or 60 min. At the cathode, significant increases of staining intensity in periosteal osteoblasts for cAMP, cGMP, and PGE were found at 15 and 60 min. At the anode, a significant rise in the staining intensity of these cells for PGE was seen at 15 min; at 60 min, cGMP and PGE, but not cAMP, were elevated. These results demonstrate the usefulness of the immunohistochemical technique in detecting relative changes in mineralized tissue cell content of cyclic nucleotides and prostaglandins in response to local application of physical stimuli of short and long duration.