Rapid desensitization of PC12 cells stimulated with high concentrations of extracellular S100

Undifferentiated PC12 cells undergo apoptosis, via a calcium-induced calcium release mechanism, when the calcium-binding protein purified from bovine brain (native S100) is present in micromolar concentration in the medium. This process begins when S100 binds to specific membrane binding sites and involves up to 50% of the cell population. In the experiments reported here, we demonstrate that, by utilizing [3H]S100, the S100 protein can be displaced from its binding sites only during the first 10 min of incubation. This fact is due to an internalization mechanism, having a time-course with a plateau after 10-20 min of incubation. The native form of S100 is a mixture of two different S100 isoforms: S100A1 (20%) and S100B (80%). Using confocal microscopy and monoclonal antibodies, we demonstrated that only one of these isoforms, S100A1, was autoexpressed in more than 50% of the PC12 cells analysed. After cell incubation with 2 microM native S100, S100B also appears in PC12 cells, with a maximum presence after 10 min of incubation. This fact seems to indicate that this isoform, at least, is effectively translocated when stimulated with external native S100. From the data reported, it is possible to hypothesize that, in PC12 cells, a possible homeostatic mechanism is present that can counteract the effect of a continuously applied lethal stimulus (stimuli) on cell viability.