Regulation of E-selectin, P-selectin, and intercellular adhesion molecule 1 expression in mouse cremaster muscle vasculature.
OBJECTIVE
To investigate the expression of P- and E-selectin and intercellular adhesion molecule 1 (ICAM-1) in the vasculature of mouse cremaster muscles both with and without tumor necrosis factor alpha (TNF-alpha) treatment.


METHODS
Mice received injections of monoclonal antibody to P-selectin, E-selectin, or ICAM-1 before fixation to restrict detection to antigen expressed on the endothelial surface. Whole-mount preparations of mouse cremaster muscles were fixed in acetone and stained using biotinylated secondary antibody and peroxidase-conjugated streptavidin.


RESULTS
P-selectin is expressed on the endothelial surface of cremaster muscle venules within 10 minutes after exteriorization. Expression increases upon treatment with TNF-alpha (2 hours), reflecting transcriptional regulation of P-selectin expression in situ. The baseline E-selectin expression is patchy and barely detectable but shows a significant upregulation after treatment with TNF-alpha. [CAM-1 is constitutively expressed in unstimulated mouse cremaster venules and slightly upregulated after 2 hours of TNF-alpha treatment. Under baseline conditions, neither E-selectin, P-selectin, nor ICAM-1 is detectable in arterioles or capillaries. After TNF-alpha treatment, arterioles stain faintly for P-selectin, but not E-selectin or ICAM-1.


CONCLUSIONS
The temporal and spatial pattern of expression of P- and E-selectin and ICAM-1 is consistent with the functional role of these molecules in mediating preferential leukocyte rolling and adhesion in mouse cremaster muscle venules.
Jung
U
U
Department of Biomedical Engineering, University of Virginia Health Sciences Center, Charlottesville 22908, USA.
Ley
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001160:Arterioles / Q000276:immunology / Q000378:metabolism; D019040:E-Selectin / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D018799:Intercellular Adhesion Molecule-1 / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000187:drug effects* / Q000601:surgery; D019007:P-Selectin / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014699:Venules / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D019040:E-Selectin; D019007:P-Selectin; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1