Substance P (SP) elicits mast cell degranulation from mucosal and connective tissue-type mast cells in rodents. We reported that SP induced degranulation from mast cells grown in stem cell factor (SCF). We now examined the production of eotaxin by SP stimulated bone marrow cultured mast cells (BMCMC). Primary cultures of murine BMCMC were maintained in either SCF (50ng/ml) or 20% Concanavalin A-stimulated spleen conditioned medium (CM) for 4-6 weeks. Specific serotonin release to SP was tested before each experiment. The mast cells were then incubated with or without 10- 4 M substance P for different times. Total RNA was isolated for RT-PCR and Northern blot analysis for eotaxin mRNA expression. Eotaxin protein expression was examined by immunocytochemistry and Western blot. BMCMC in SCF, but not BMCMC in CM, released serotonin upon challenge with SP. BMCMC in SCF constituitively expressed eotaxin mRNA and the level was markedly increased 30 mins after stimulation with SP. Eotaxin protein was demonstrable by immunocytochemistry or Western blot in BMCMC in SCF after stimulation with SP. By contrast, eotaxin protein was not detected in BMCMC in SCF stimulated by IgE and antigen or in BMCMC in CM by either SP or IgE and antigen. Our data demonstrate that the peptide SP stimulates eotaxin production from BM-CMC derived in SCF and suggests that SP-induced eotaxin production involves a signal transduction pathway distinct from signaling via Fc εRI.
|Original language||English (US)|
|State||Published - Mar 20 1998|
ASJC Scopus subject areas
- Molecular Biology