Dataset Information
An involvement of neurokinin-1 receptor in Fc?R?-mediated RBL-2H3 mast cell activation.
Ontology highlight
ABSTRACT: Objective and design
To determine whether the neurokinin-1 receptor (NK1R) plays a role in the activation of RBL-2H3 mast cells after Fc?R? aggregation.Materials and methods
NK1R expression in RBL-2H3 cells was inhibited by small hairpin RNA (shRNA) against NK1R, and determined by western blotting. For activation, both NK1R knockdown and control RBL-2H3 cells were sensitized by dinitrophenol (DNP)-specific IgE and stimulated with the antigen DNP-bovine serum albumin (BSA). Following the activation of RBL-2H3 cells, monocyte chemoattractant protein (MCP-1) production and intracellular calcium flux were monitored by ELISA and confocal microscopy assay, respectively. For investigation of the signaling mechanism, phosphorylation of mitogen-activated protein kinases (MAPKs) after RBL-2H3 cell activation was assessed by western blotting.Results
shRNA-NK1R mediated an effective inhibition of NK1R expression in RBL-2H3 cells. Protein production of MCP-1 was reduced by more than 55 % in NK1R knockdown RBL-2H3 cells compared with control RBL-2H3 cells. In addition, both calcium mobilization and phosphorylation levels of MAPKs (Erk1/2, JNK, and p38) after DNP-BSA stimulation (via Fc?R?) were decreased due to the inhibition of NK1R expression.Conclusion
NK1R is required for the activation of RBL-2H3 cells following Fc?R? engagement and involved in the regulation of MAPK signaling pathways.
SUBMITTER: Fang X
PROVIDER: S-EPMC3472057 | biostudies-literature | 2012 Nov
REPOSITORIES: biostudies-literature
Publications
An involvement of neurokinin-1 receptor in FcεRΙ-mediated RBL-2H3 mast cell activation.
Inflammation research : official journal of the European Histamine Research Society ... [et al.] 20120721 11
<h4>Objective and design</h4>To determine whether the neurokinin-1 receptor (NK1R) plays a role in the activation of RBL-2H3 mast cells after FcεRΙ aggregation.<h4>Materials and methods</h4>NK1R expression in RBL-2H3 cells was inhibited by small hairpin RNA (shRNA) against NK1R, and determined by western blotting. For activation, both NK1R knockdown and control RBL-2H3 cells were sensitized by dinitrophenol (DNP)-specific IgE and stimulated with the antigen DNP-bovine serum albumin (BSA). Follow ...[more]
