Identification of Nepsilon-(carboxymethyl)lysine-positive cells in astroglia-rich primary cultures.
Astroglia-rich primary cultures from rat brain were used to investigate the presence in glial cells of Nepsilon-(carboxymethyl)lysine (CML), an advanced glycation endproduct. Westernblot analysis of homogenates of rat brain as well as of astroglia-rich cultures demonstrated the presence of CML-modified proteins in these samples. Immunocytochemical staining of astroglia-rich cultures revealed that only a minority of the cells in these cultures were intensively stained for CML. The staining intensity of CML-positive cells was strongly reduced, if the cells were not permeabilized, indicating that intracellular proteins were CML-modified. The CML-positive cells were identified as astrocytes and oligodendrocytes by double-labelling immunocytochemical staining for CML and the cellular markers galactocerebroside, myelin basic protein and glial fibrillary acidic protein. In contrast to other glial cells, microglial cells in astroglia-rich cultures were CML-negative. The finding that only a minority of cells in astroglia-rich cultures contains high amounts of intracellular CML-modified proteins indicates that specific properties of these CML-positive cells are responsible for the CML-formation in these cells.
Pawlowski
P G
PG
Physiologisch-chemisches Institut, Universität Tübingen, Germany.
Dringen
R
R
D000818:Animals; D001253:Astrocytes / Q000737:chemistry* / Q000378:metabolism; D015153:Blotting, Western; D001923:Brain Chemistry; D018929:Cell Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D017127:Glycation End Products, Advanced / Q000032:analysis* / Q000378:metabolism; D007150:Immunohistochemistry; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D017127:Glycation End Products, Advanced; C048496:N(6)-carboxymethyllysine; D008239:Lysine