Nuclear and cytosolic calcium changes in osteoclasts stimulated with ATP and integrin-binding peptide.
Cytosolic calcium modulates the activity of osteoclasts, large multinucleate cells that resorb bone. Nuclear events, such as gene transcription, are also calcium-regulated in these cells, and fluorescence imaging has suggested that calcium signals produced by some stimuli are specifically targeted to, or amplified within, osteoclast nuclei. We used two alternative techniques of dye loading to examine the changes of intracellular calcium induced in rat osteoclasts by three stimuli. Osteoclasts loaded with the calcium indicator Fura-2 by the acetoxymethyl (AM) ester technique appeared to display marked nuclear calcium amplification. During stimulation with integrin-binding peptides, ATP, or high extracellular calcium, fluorescence ratios recorded from the nuclei rose higher than did ratios recorded from extranuclear regions. In contrast, nuclear calcium amplification was not observed after AM loading in the presence of the anion transport inhibitor sulfinpyrazone, nor in osteoclasts injected with Fura-2 conjugated to a high MW dextran. In these cells, nuclear fluorescence ratios were equal to the extranuclear values at all times: upon stimulation by an agonist, the nuclear and cytosolic calcium concentrations increased by the same amount. The calcium changes seen in stimulated osteoclasts can no longer be taken as evidence for the general validity of the phenomenon of nuclear calcium amplification.
Parkinson
N
N
Department of Physiology, University College London, UK. nicola.parkinson@ucl.ac.uk
Bolsover
S
S
Mason
W
W
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000032:analysis* / Q000378:metabolism*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D003911:Dextrans / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D016257:Fura-2 / Q000737:chemistry / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D016023:Integrins / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology*; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D013050:Spectrometry, Fluorescence; D013442:Sulfinpyrazone / Q000737:chemistry / Q000494:pharmacology
D003911:Dextrans; D005456:Fluorescent Dyes; D016023:Integrins; D009842:Oligopeptides; D000255:Adenosine Triphosphate; C049554:glycyl-arginyl-glycyl-aspartyl-serine; D002118:Calcium; D016257:Fura-2; D013442:Sulfinpyrazone