Prostaglandin D2 generation in mouse bone marrow-derived mast cells exposed to dexamethasone is associated with endogenous peroxidase activity.
The appearance of fixative-sensitive peroxidase activity in the nuclear envelope and endoplasmic reticulum of bone marrow-derived mast cells (BMMC) cultured in the presence of 1 microM dexamethasone (DM) for up to 14 days and its relationship with immunologic release of prostaglandin D2 (PGD2) by these cells were studied. Endogenous peroxidase activity, previously shown as a marker of arachidonic acid metabolism in various cell types, was visualized by cell incubation in 3,3' diaminobenzidine-containing solution before glutaraldehyde fixation. PGD2 release was induced by passive sensitization of BMMC with an optimal dose of monoclonal IgE and subsequent challenge with specific a antigen. We found that 4-week-old BMMC, used as the starting population of the present study, exhibited immature morphologic features, did not present peroxidase activity when cytochemically processed, and released minute amounts of PGD2 in response to IgE-dependent stimulation. When such BMMC were exposed to DM during 24 hours, they showed aldehyde-inhibited peroxidase activity in the perinuclear envelope and a few endoplasmic reticulum segments. As compared with untreated cells, 24-hour DM-exposed BMMC released higher amounts of PGD2 upon immunologic stimulation. After an additional 14-day period of DM exposure, an intense peroxidase activity was detected in the perinuclear envelope and the endoplasmic reticulum of BMMC, which, under immunologic stimulation, released as much as 42.4 +/- 14.7 ng of PGD2/1 x 10(6) cells. Aminotriazole (20 and 50 mM) extinguished both peroxidase activity and PGD2 release from BMMC whereas indomethacin (1 microM) suppressed PGD2 production, but did not alter endogenous peroxidase activity. Previous cell fixation with glutaraldehyde totally inhibited endogenous peroxidase reaction in DM-exposed BMMC. Moreover, 14-day DM-exposed BMMC exhibited morphologic characteristics of mature mast cells and possessed alcian blue+/safranin+ granules. Therefore, the present data suggest that appearance of peroxidase activity in the nuclear envelope and the endoplasmic reticulum of DM-exposed BMMC is associated with the ability of the cells to synthetize PGD2 and appears as a cytochemical marker of the in vitro maturation of mouse bone marrow-derived mast cells.
Michel
L
L
INSERM U312, Laboratoire de Dermatologie, Hôpital Henri Mondor, Creteil, France.
Pitton
C
C
Prost
C
C
Massé
J M
JM
Mencia-Huerta
J M
JM
Burtin
C
C
Maclouf
J
J
Breton-Gorius
J
J
Benveniste
J
J
Dubertret
L
L
D000447:Aldehydes / Q000494:pharmacology; D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D001386:Azides / Q000494:pharmacology; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D003486:Cyanides / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology*; D006636:Histamine Release / Q000187:drug effects; D006651:Histocytochemistry; D007213:Indomethacin / Q000494:pharmacology; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010544:Peroxidases / Q000378:metabolism; D015230:Prostaglandin D2 / Q000096:biosynthesis*
D000447:Aldehydes; D001386:Azides; D003486:Cyanides; D003907:Dexamethasone; D010544:Peroxidases; D015230:Prostaglandin D2; D007213:Indomethacin; D000640:Amitrole