关键词:
Mechanical stimulation
Inositol (1,4,5)-trisphosphate
Ca2+ response
FRET
Fluorescent biosensor
Spatial dynamics
摘要:
In the present study, the contribution of inositol (1,4,5)-trisphosphate [Ins(1,4,5)P-3] generation on the mechanical-stimulation-induced Ca2+ response was investigated in HSY-EA1 cells. Mechanical stimulation induced a local increase in the cytosolic concentration of Ins(1,4,5)P-3 ([IP3](i)), as indicated by the Ins(1,4,5)P-3 biosensor LIBRAvIII. The area of this increase expanded like an intracellular Ins(1,4,5)P-3 wave as [IP3](i) increased in the stimulated region. A small transient [IP3](i) increase was subsequently seen in neighboring cells. The phospholipase C inhibitor U-73122 abolished these Ins(1,4,5)P-3 responses and resultant Ca2+ releases. The purinergic receptor blocker suramin completely blocked increases in [IP3](i) and the Ca2+ release in neighboring cells, but failed to attenuate the responses in mechanically stimulated cells. These results indicate that generation of Ins(1,4,5)P-3 in response to mechanical stimulation is primarily independent of extracellular ATP. The speed of the mechanical-stimulation-induced [IP3](i) increase was much more rapid than that induced by a supramaximal concentration of ATP (1 mM). The contribution of the Ins(1,4,5)P-3-induced Ca2+ release was larger than that of Ca2+ entry in the Ca2+ response to mechanical stimulation in HSY-EA1 cells.