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Journal of Histochemistry and Cytochemistry, Vol. 45, 1379-1392, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

Immunocytochemical Localization of Chymase to Cytoplasmic Vesicles After Rat Peritoneal Mast Cell Stimulation by Compound 48/80

Gary R. Logina,b,c, Mikako Aokid, Midori Yamakawac, Laurelúcia O. Lunardie, Eleni C. Digenisc, Naoko Tandac, Lawrence B. Schwartzf, and Ann M. Dvorakb,c
a Department of Pathology, Harvard School of Dental Medicine, Boston, Massachusetts
b Department of Pathology, Harvard Medical School, Boston, Massachusetts
c Department of Pathology, Beth Israel-Deaconess Medical Center, Boston, Massachusetts
d Department of Dermatology, Nippon Medical School, Tokyo, Japan
e Department of Morphology, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, São Paulo, Brazil
f Department of Medicine, Virginia Commonwealth University, Richmond, Virginia

Correspondence to: Gary R. Login, Dept. of Pathology, Beth Israel Hospital, 330 Brookline Ave., Boston, MA 02215.

The subcellular events responsible for release of mediators by mast cells may help to clarify roles for mast cells in health and disease. In this study we show that the granule-associated protease chymase is also within cytoplasmic vesicles in appropriately stimulated rat peritoneal mast cells. Rat peritoneal mast cells were recovered before or 1-10 sec after exposure to the secretogogue compound 48/80 (10 µg/ml) and then were examined by radioimmunoassay to quantify histamine release or were processed, using routine methods for postembedding immunoelectron microscopy, to identify the subcellular localization of chymase. In comparison to unstimulated cells, compound 48/80 stimulated cells in two independent experiments showed an increase (15%, 28%) in the surface area of the cell and a decrease (12%, 6%) in the surface area of the total granule compartment before degranulation channel formation. These global cellular changes occurred in a background of transient but significant (p<0.01) increases in the area and number of chymase-immunoreactive vesicles per µm2 cytoplasm. These changes were detectable at 5 or 7 sec after stimulation with compound 48/80 but returned to near prestimulation levels by 9 or 10 sec after addition of compound 48/80 (total cumulative histamine release was 28% by 8 sec and 47% by 14 sec). These observations suggest that vesicles participate in the early stages of regulated secretion of chymase from rat peritoneal mast cells. (J Histochem Cytochem 45:1379-1391, 1997)

Key Words: rat, vesicular transport, secretion, morphometry, electron microscopy, immunocytochemistry, mast cell, chymase


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