Leukocytes express connexin 43 after activation with lipopolysaccharide and appear to form gap junctions with endothelial cells after ischemia-reperfusion.
Levels and subcellular distribution of connexin 43 (Cx43), a gap junction protein, were studied in hamster leukocytes before and after activation with endotoxin (lipopolysaccharide, LPS) both in vitro and in vivo. Untreated leukocytes did not express Cx43. However, Cx43 was clearly detectable by indirect immunofluorescence in cells treated in vitro with LPS (1 micrograms/ml, 3 hr). Cx43 was also detected in leukocytes obtained from the peritoneal cavity 5-7 days after LPS-induced inflammation. In some leukocytes that formed clusters Cx43 immunoreactivity was present at appositional membranes, suggesting formation of homotypic gap junctions. In cell homogenates of activated peritoneal macrophages, Cx43, detected by Western blot analysis, was mostly unphosphorylated. A second in vivo inflammatory condition studied was that induced by ischemia-reperfusion of the hamster cheek pouch. In this system, leukocytes that adhered to venular endothelial cells after 1 hr of ischemia, followed by 1 hr of reperfusion, expressed Cx43. Electron microscope observations revealed small close appositions, putative gap junctions, at leukocyte-endothelial cell and leukocyte-leukocyte contacts. These results indicate that the expression of Cx43 can be induced in leukocytes during an inflammatory response which might allow for heterotypic or homotypic intercellular gap junctional communication. Gap junctions may play a role in leukocyte extravasation.
Jara
P I
PI
Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago.
Boric
M P
MP
Sáez
J C
JC
D000818:Animals; D002455:Cell Division; D018031:Connexin 43 / Q000096:biosynthesis* / Q000097:blood; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000502:physiology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D017629:Gap Junctions / Q000648:ultrastructure*; D015151:Immunoblotting; D007962:Leukocytes / Q000502:physiology* / Q000648:ultrastructure; D008070:Lipopolysaccharides; D008297:Male; D008647:Mesocricetus; D010538:Peritonitis / Q000378:metabolism*; D015427:Reperfusion Injury; D014699:Venules / Q000473:pathology
D018031:Connexin 43; D008070:Lipopolysaccharides