Nuclear versus perinuclear and cytoplasmic calcium in osteoclasts

We measured fluorescence from the calcium indicator Fluo-3 in multinucleated osteoclasts. In the initial state, each nucleus is surrounded by a ring of bright fluorescence. Following activation of purinergic receptors by 100 microM ATP there is a pulse of cellular fluorescence increase, and nuclear fluorescence intensity becomes greater than that of the cytoplasm. This is followed by a period during which the fluorescence of the cell decreases below that of the initial state. During the pulsed increase following purinergic receptor activation, the perinuclear fluorescence intensity does not increase as much as that in the nuclear centers and, following this pulse, the perinuclear fluorescence intensity decreases more than that in the nuclear centers, relative to the initial state. Measurements in which Mn2+ was introduced into the cell show that the number of Fluo-3 molecules per unit horizontal area in the nuclear centers is slightly greater than that in the perinuclear regions, and more than twice that in the surrounding cytoplasm. These results show that there is a much higher free calcium concentration in the perinuclear regions than in the nuclear centers in the initial state, with a release of free calcium from the perinuclear regions following activation of the purinergic receptors. These data also provide evidence that the free calcium concentration in the nuclear centers is lower than in the cytoplasm in the initial state.