Inhibition of degranulation in mast cells attached to a hydrogel through defective microtubule tracts.

Mast cells (MCs) are important effectors of the immediate allergic response. MCs are distributed throughout various tissues and organs, and adhere to extracellular matrix (ECM) with broad stiffness in the body. Here we compared cellular responses following antigen stimulation in MCs on glass-base dishes with and without a hydrogel. We found that an antigen-induced increase in intracellular Ca2+ concentration was suppressed slightly in cells on hydrogel-coated dishes compared with those on non-coated dishes, whereas their subsequent degranulation was largely inhibited in cells adherent to the hydrogel. Focusing on focal adhesions (FAs), vinculin was distributed in a dot-like manner at the bottom of resting cells on non-coated dishes but not on hydrogel-coated dishes. According to antigen stimulation, phosphorylation of focal adhesion kinase and additive vinculin accumulation to FAs were promoted in cells on non-coated dishes, but were diminished on hydrogel-coated dishes. Moreover, microtubule reorganization and acetylation (which have important roles in MC degranulation) were also suppressed in activated MCs adherent to the hydrogel. These findings suggest that adhesion to a hydrogel led to failure of composition of functional FAs and microtubule tracts, which resulted in suppression of MC degranulation following antigen stimulation.