Tumor necrosis factor alpha immunoreactivity of rat peritoneal mast cell granules decreases during early secretion induced by compound 48/80: An ultrastructural immunogold morphometric analysis

被引:23
作者
Beil, WJ
Login, GR
Aoki, M
Lunardi, LO
Morgan, ES
Galli, SJ
Dvorak, AM
机构
[1] BETH ISRAEL HOSP,DEPT PATHOL,BOSTON,MA 02215
[2] HARVARD UNIV,SCH MED,BOSTON,MA
[3] HARVARD UNIV,SCH DENT MED,BOSTON,MA 02115
[4] NIPPON MED COLL,DEPT DERMATOL,TOKYO,JAPAN
[5] UNIV SAO PAULO,FAC MED RIBEIRAO,DEPT MORPHOL,BR-05508 SAO PAULO,BRAZIL
关键词
tumor necrosis factor-alpha; mast cell; granules; secretion; compound; 48/80; microwave fixation; ultrastructure; immunogold; morphometrics;
D O I
10.1159/000237267
中图分类号
R392 [医学免疫学];
学科分类号
100102 ;
摘要
We used fast (seconds) and ultrafast (milliseconds) microwave energy-assisted chemical fixation protocols, postembedding immunogold staining, and a morphometric analysis to investigate the early morphological changes and the TNF-alpha immunoreactivity in the cytoplasmic granules of rat peritoneal mast cells that had been stimulated to secrete by exposure to compound 48/80, Exposure to compound 48/80 induced the development of increased numbers of cytoplasmic granules that exhibited decreased electron density; these granules often also appeared swollen. These granule alterations were accompanied by a significantly decreased proportion of granules that were positive for TNF-alpha immunoreactivity. We also calculated the density of TNF-alpha labeling/mu(2) in both dense (unaltered) and altered granules in specimens. TNF-alpha immunoreactivity was present in dense granules (regardless of whether or not the specimens had been stimulated with compound 48/80) and in cells that were fixed with either fast or ultrafast microwave energy. However, altered granules exhibited a decreased density of TNF-alpha label. These findings show that changes in the immunolocalization and/or density of TNF-alpha immunoreactivity occur very rapidly upon stimulation of rat peritoneal mast cells with compound 48/80.
引用
收藏
页码:383 / 389
页数:7
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