Heterogeneity of intracellular calcium responses to parathyroid hormone and thrombin in primary osteoblast-like cells and UMR106-01 cells: Correlations with culture conditions, intracellular calcium concentration and differentiation state

Abstract

The present study evaluates the effect of parathyroid hormone (PTH) on intracellular calcium. Intracellular calcium ion concentrations ([Ca 2+] i) in fetal rat osteoblasts in primary culture (ROB) and in UMR106-01 osteogenic sarcoma cells were monitored as changes in the ratio (R) of Fura-2 fluorescence intensities in single cells as well as populations of cells. In both single ROB and UMR106-01 cells, addition of 10 −7 M rat PTH1–34 and 3 NIH units/ml human thrombin resulted in heterogeneous responses in R values and therefore [Ca 2+] i. PTH-induced calcium responsiveness of ROB was dependent on culture conditions, such that response frequencies were positively correlated with the percentage of fetal calf serum in the culture medium. PTH responsive ROB and UMR106-01 cells had significantly higher resting [Ca 2+] i than unresponsive cells. PTH- or thrombin-mediated calcium signalling appeared not to be correlated to alkaline phosphatase activity in single ROB. Low percentages of cells responded to PTH in comparison to thrombin suggesting that an increase in [Ca 2+] i is not a common PTH signalling pathway in osteoblasts in primary culture. Our data suggest that activation of this signalling pathway by PTH is culture condition dependent, possibly via a cell-cycle related increase in sensitivity of the pathway.