1. Fluorescence Measurements of [Ca2+]I after shear stress of RBL cells
Effects of Shear Stress on Intracellular Calcium Change and Histamine Release in Rat Basophilic Leukemia (RBL-2H3) Cells
Fei Wei, WenzhongYang, Luwei Zhou
Phys. Dept., Fudan Univ., Shanghai, People’s Republic of China
Abstract:
A mast cell RBL-2H3 was used to study cellular responses to shear stress generated by a rotating rotor in a cell dish. The intracellular calcium was studied by confocal fluorescence microscopy with Fluo-3/AM staining and the released histamine was measured with a fluorescence spectrometer using o-phthalaldehyde (OPA ) staining. An elevation of [Ca2+] occurred immediately after the shear stress, followed by histamine release. Furthermore, Ruthenium red, a transient receptor potential vanilloid (TRPV) inhibitor, could effectively block the shear stress–induced histamine release, suggesting that TRPV membrane proteins are the likely targets of the shear stress.
Fluorescence Measurements of [Ca2+]I after shear stress of RBL cells
1.Stained with fluo-3/AM
2.Confocal fluorescence microscopy
3.Ca2+ fluorescence intensities in a cell in the Ca 2+free saline:
η-viscosity
ω-rotation speed
Shear  [Ca2+ ]i
Histamine detecting method
TRPV4 channel
OPA邻苯二甲醛+组胺
Cell viability
Shear  histamine
Shear  TRPV Ca2+ & histamine
Conclusion :
The change of [Ca2+ ]i and then histamine release can be induced by shear stress in RBL-2H3 cells . We speculate the TRPV4 channel involves in the response of RBL-2H3 to the physical stimuli. We would study the response of RBL cells to reciprocating force.
Reference:
[1] Wenzhong Yang et al. Effects of Shear Stress on Intracellular Calcium Change and Histamine Release in Rat Basophilic Leukemia (RBL-2H3) Cells, Journal of Environmental Pathology, Toxicology and Oncology, 28(3):223–230 (2009)
[2] Wen-Zhong Yang et al. Effects of Low Power Laser Irradiation on Intracellular Calcium and Histamine Release in RBL-2H3 Mast Cells, Photochemistry and Photobiology, 2007, 83: 979–984