Abstract
Using laser confocal microscopy and a Ca 2+ indicator, fluo-3, we find that osteoclasts respond to IL-1α with a sustained increase (4 to 40 min) in cytosolic free calcium ([Ca 2+] i). The delay time ( T 1/2) varies from 2 to 8 minutes. These effects cannot be mimicked by application of cAMP and can be blocked by removal of Ca 2+ from the bathing medium. Pre-incubation with indomethacin (30 μM) does not block the IL-1α effect on [Ca 2+] i. We employ a ′micropipette′ method and a ′pieces of cover slip′ method to isolate single osteoclasts. Our results provide evidence that IL-1α directly stimulates osteoclasts via a [Ca 2+] i signalling pathway. We postulate that this slowly developing, long-lasting increase in [Ca 2+] i in osteoclasts may account for the stimulatory effect of IL-1α on osteoclastic bone resorption in conjunction with signalling molecules from osteoblasts.
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