Orai1 regulates intracellular calcium, arrest, and shape polarization during neutrophil recruitment in shear flow.
Orai1 was reported to function as a calcium channel subunit that facilitates store operated calcium entry (SOCE) in T cells and is necessary for formation of the immune synapse. We reasoned that SOCE via Orai1 might regulate PMNs activation during recruitment to inflamed endothelium. Orai1 function was assessed by real-time imaging of calcium transients as PMNs were stimulated to roll, arrest, and migrate on E-selectin and ICAM-1 in shear flow. Calcium entry was significantly reduced when Orai1 function was impaired by heterozygous knockout in a mouse model or by siRNA knockdown in HL-60 cells. Reduced Orai-1 expression correlated with the delayed onset of arrest and reduced ability to transition to a polarized migratory phenotype. Inhibition of SOCE by treatment with 2-APB, or blocking phospholipase C (PLC) mediated calcium store release with U73122, abrogated formyl peptide induced calcium elevation, and delayed subsequent cell arrest and polarization. These results suggest that calcium entry via Orai1 is the predominant SOCE that cooperates with cytoplasmic calcium store release in coordinating integrin-dependent PMN arrest and migration in the acute response to inflammation.
Schaff
Ulrich Y
UY
Department of Biomedical Engineering, University of California, Davis, CA, USA.
Dixit
Neha
N
Procyk
Emily
E
Yamayoshi
Itsukyo
I
Tse
Tiffany
T
Simon
Scott I
SI
D000818:Animals; D001775:Blood Circulation / Q000502:physiology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics / Q000502:physiology*; D002465:Cell Movement / Q000235:genetics* / Q000502:physiology; D016764:Cell Polarity / Q000235:genetics*; D049109:Cell Proliferation; D048430:Cell Shape / Q000235:genetics; D002478:Cells, Cultured; D018922:HL-60 Cells; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D009504:Neutrophils / Q000378:metabolism / Q000473:pathology / Q000502:physiology*; D000071740:ORAI1 Protein; D033081:Shear Strength / Q000502:physiology
D015220:Calcium Channels; D000071740:ORAI1 Protein; C510746:ORAI1 protein, human; D002118:Calcium