The effect of human calcitonin on the cytoplasmic spreading of rat osteoclasts.

We previously found that calcitonin (CT), which inhibits osteoclastic bone resorption, abolished the cytoplasmic motility of isolated osteoclasts. The transition from motility to immotility was accompanied by a characteristic change in cell shape which coincided with the loss of pseudopodial ruffling activity and gradual cytoplasmic retraction. In this report we used computer-assisted morphometry to quantify the reduction in cytoplasmic spreading induced by human CT (hCT). Osteoclasts, isolated from neonatal rat long bones, were allowed to spread on a plastic surface for 120 min. The outlines of six osteoclasts were recorded on a time-lapse video recorder. hCT, other hormones, or plasma samples were then added. The outlines of the same six osteoclasts were recorded after incubation, and the area covered by the cells after incubation was calculated as a percentage of the area covered by the same cells before hormone addition. Osteoclastic spreading was reduced by hCT in a dose-dependent manner and was significantly decreased by concentrations of 2 pg/ml and above. High concentrations of PTH and 1,25-dihydroxyvitamin D3 had no effect on osteoclasts. This technique is a sensitive and quantitative assay of hCT concentration which is unaffected by the presence of other calcium-regulating hormones likely to be present in plasma.