Luminal calcium regulates the inositol trisphosphate receptor of rat basophilic leukemia cells at a cytosolic site.
Hormones, growth factors, and other stimuli can generate Ca2+ spikes and waves by activation of the phosphoinositide (PI) pathway. The sources of these Ca2+ signals are inositol 1,4,5-trisphosphate (IP3)-dependent Ca2+ stores. Here we use a rapid perfusion apparatus to measure the release of 45Ca2+ from permeabilized rat basophilic leukemia (RBL) cells to investigate the regulation of IP3-mediated Ca2+ release by cytosolic and luminal Ca2+. At 200 nM IP3, Ca2+ release was potentiated by an increase in the cytosolic Ca2+ concentration. This potentiation by Ca2+ was nearly absent at 500 nM IP3. Previous studies in smooth muscle cells and neurons showed an inhibition of Ca2+ release above 300 nM Ca2+. In contrast, no such inhibition was observed in RBL cells. When assayed at low cytosolic Ca2+ concentrations, IP3-mediated release was steeply dependent upon luminal Ca2+ concentration. At high luminal Ca2+ concentration, 1 microM IP3 released most of the stored Ca2+ even in the complete absence of cytosolic Ca2+. However, at high cytosolic Ca2+ concentrations (890 nM), IP3-mediated release was no longer steeply dependent upon the luminal Ca2+ concentration. Furthermore, high concentrations of BAPTA inhibited IP3-mediated release in the absence of cytosolic Ca2+. This suggests that a rapid and local luminal Ca2+ feedback is generated by luminal Ca2+ ions binding to cytosolic sites of the same channel or closely associated channels. This "luminal Ca2+ feedback" can be initiated by an increase in the concentration either of IP3, of cytosolic Ca2+, or of luminal Ca2+. It is likely that "luminal Ca2+ feedback" is exploited by cells in both the initiation and termination of Ca2+ spikes.
Horne
J H
JH
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Meyer
T
T
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D007202:Indicators and Reagents; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D053496:Inositol 1,4,5-Trisphosphate Receptors; D015471:Leukemia, Basophilic, Acute; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D015220:Calcium Channels; D007202:Indicators and Reagents; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018160:Receptors, Cytoplasmic and Nuclear; D004533:Egtazic Acid; D015544:Inositol 1,4,5-Trisphosphate; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium