Related Experiment Videos
[A study on the changes of calcium signal in mast cells activated by antigen]
1Department of Respiratory Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
Objective:
To study the relationship between the calcium signal and activation of mast cells, laying the foundation for discovering a new potential theraputical target of signal transduction for type I anaphylactic diseases like asthma.
Methods:
The fluorescent Ca2+ indicator Fluo-3 was used to observe and quantitate the calcium signal directly in activated RBL-2H3 mast cells by a flow cytometer and a confocal fluorescence microscope with laser.
Results:
Antigens elicited significant increase in intracellular Ca2+ concentration ([Ca2+]i) and histamine release in sensitized cells, maximal [Ca2+]i increase being reached (525 +/- 42) nmol/L (n = 12) within 90-110 s after stimulation. Increase of fluo-3-fluorescence intensity was also observed with the confocal fluorescence microscope in cytosol. No significant increase of [Ca2+]i was observed after addition of EDTA in sensitized cells from an initial value of (79 +/- 3) nmol/L, to (80 +/- 4) nmol/L (P > 0.05, n = 12), but tyrosine phosphorylation was readily observed, and the indirect fluorescent intensity was significantly greater than that in the control group. No significant change of [Ca2+]i was observed with the addition of DNP-BSA after PMA (P > 0.05, n = 8).
Conclusion:
Calcium is involved in the activation process of mast cells stimulated by antigens.