Loss of TRPC1-mediated Ca2+ influx contributes to impaired degranulation in Fyn-deficient mouse bone marrow-derived mast cells.
MC degranulation requires the influx of calcium from the extracellular environment. Orai1/STIM1 is essential to MC SOCE, as shown in rat peritoneal MCs, the rat MC lines (RBL-2H3), or in Orai1 null embryo liver-derived, cultured MCs. However, minimal information exists about the role of other calcium channels expressed on these cells. Here, we demonstrate that the nonselective TRPC1 participates in FcεRI-mediated calcium entry in mouse BMMCs. We found that Fyn null MCs, which have an impaired degranulation response, expressed reduced levels of TRPC1, had normal depletion of intracellular calcium stores but an impaired calcium influx, and failed to depolymerize cortical F-actin (a key step for granule-plasma membrane fusion). Partial RNAi silencing of TRPC1 expression in WT MCs (to the level of Fyn null MCs) mimicked the Fyn null defect in calcium influx, cortical F-actin depolymerization, and MC degranulation. Ectopic expression of Fyn or TRPC1 in Fyn null MCs restored calcium responses and cortical F-actin depolymerization and increased MC degranulation. Together with our findings that expression of Orai1 is not altered in Fyn null MCs, our findings suggest that TRPC1 participates in calcium influx and other key events required for MC degranulation. This demonstrates that in addition to a role described previously for Orai1 in promoting MC degranulation, nonselective cation channels participate in promoting the exocytotic response.
Suzuki
Ryo
R
National Institutes of Health, Bethesda, MD 20892-1930, USA.
Liu
Xibao
X
Olivera
Ana
A
Aguiniga
Lizath
L
Yamashita
Yumi
Y
Blank
Ulrich
U
Ambudkar
Indu
I
Rivera
Juan
J
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001853:Bone Marrow / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics; D008407:Mast Cells / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D051076:Proto-Oncogene Proteins c-fyn / Q000172:deficiency* / Q000235:genetics; D051381:Rats; D050052:TRPC Cation Channels / Q000502:physiology*; D019284:Thapsigargin / Q000494:pharmacology; D019061:src-Family Kinases / Q000172:deficiency
D000199:Actins; D004791:Enzyme Inhibitors; D008164:Luminescent Proteins; D050052:TRPC Cation Channels; C096351:transient receptor potential cation channel, subfamily C, member 1; D019284:Thapsigargin; C494960:Fyn protein, mouse; D051076:Proto-Oncogene Proteins c-fyn; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D002118:Calcium