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Shear stress‐sensitive TRP channels modulate calcium influx in activated T cells
Author(s) -
Holtzclaw J. David,
Brown Rachel C.,
O’Neil Roger G.
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1349
Subject(s) - trpv , chemistry , jurkat cells , calcium , transient receptor potential channel , biophysics , trpc , voltage dependent calcium channel , mechanosensitive channels , trpc5 , calcium in biology , calcium channel , microbiology and biotechnology , ion channel , biochemistry , t cell , biology , immunology , receptor , organic chemistry , immune system , trpv1
Mechanosensitve calcium channels have been implicated in early stage lymphocyte activation. The potential role of TRP channels in regulating shear stress‐sensitive calcium influx was assess in activated Jurkat cells by calcium imaging using fura 2 ratiometric fluorescence analysis. METHODS: Jurkat cells (clone E6‐10) were activated by addition of either 100 ng/ml PMA plus 1 μg/ml A23187 or 2.5 μg/ml OKT3 (anti‐CD3) for three hours at 37 °C. Cells were washed (Hank’s balance Salt Solution), loaded with fura 2, and allowed to adhere to polyethylenimine‐coated coverslips. The coverslips were attached to a parallel plate flow chamber on an imaging workstation (Intracellular Imaging, Inc.) at 37 °C. RESULTS: Application of shear stress (3–20 dyne/cm 2 ) induced calcium influx in both anti‐CD3‐ and PMA plus A23187‐activated cells, but not in non‐activated cells. Hypotonic swelling had little influence. The shear stress‐induced calcium influx in activated cells was inhibited by antagonists of TRPV and TRPC channels (ruthenium red, SKF 96365), but not by nifedipine (L‐type channel inhibitor) or Na + ‐free buffers (TRPM4 inhibition). CONCLUSIONS: It is concluded that activated Jurkat cells express shear stress‐sensitive calcium influx and that this influx is mediated by mechanosensitive TRPV channels, and potentially TRPC channels, but not by L‐type or TRPM4 channels. (NIH Grant DK070950)