Ultrastructural study of mast cells stimulated with compound 48/80 as revealed by quick-freezing method

The ultrastructure of mast cells stimulated with compound 48/80 was examined by quick-freezing and deep-etching (QF-DE) or freeze-substitution (QF-FS) methods. Peritoneal cells including mast cells of adult male rats were stimulated in vitro with compound 48/80 at 17 degrees C for 0, 10, 30, 60 or 180 s. The QF-DE replicas revealed that the mast cells stimulated with compound 48/80 for 30 s decreased filamentous actin around secretory granules. In the QF-FS specimens, perigranular membranes in mast cells stimulated for 60 s formed pentalaminar structures between adjacent granules in their cytoplasm prior to degranulation. These findings suggest that preparatory states for degranulation occur in the whole cytoplasm of stimulated mast cells at early stages. Moreover, both QF-FS specimens and QF-DE replicas revealed a compact morphological appearance of discharged granules in the extracellular space, indicating the existence of considerable content within the granules. Skeletal structures in the granules were also demonstrated on QF-DE replicas prepared after extracting soluble elements from the cytoplasm. It is suggested that the granular contents associated with the skeletal structures are gradually detached from the discharged granules to ensure local concentration in the tissues.