Selective but nonspecific immunolabeling of enamel protein-associated compartments by a monoclonal antibody against vimentin.
Vimentin, an intermediate filament component, has been identified in many mesenchymal cells by a variety of LM and EM immunolabeling techniques. In our study, several tissue-processing conditions and monoclonal and polyclonal antibodies against vimentin were screened for immunostaining of rat incisor odontoblasts. Using postembedding colloidal gold immunocytochemistry, we were unable to detect any convincing vimentin antigenicity in these cells, but one of the monoclonal antibodies (V9-S) unexpectedly resulted in intense labeling over intra- and extracellular compartments that normally are strongly immunoreactive with anti-amelogenin antibodies. Blocking experiments showed that V9-S binding was competed by anti-amelogenin antibody. Immunoblots indicated that enamel proteins reacted with this anti-vimentin antibody after fixation with glutaraldehyde. These data suggest that the observed immunoreaction is directed against an epitope apparently created by crosslinking of enamel proteins during fixation. Although the labeling cannot be considered specific, it is nevertheless selective because it is very precisely localized over compartments containing enamel proteins and shows no binding to other calcified dental tissues, including dentin and bone. The V9-S antibody can therefore be used as a reliable probe to identify the presence and distribution of amelogenins in fixed tissues. (J Histochem Cytochem 47:1237-1245, 1999)
Josephsen
K
K
Department of Dental Pathology, Operative Dentistry and Endodontics, Royal Dental College, Faculty of Health Sciences, University of Aarhus, Aarhus, Denmark.
Smith
C E
CE
Nanci
A
A
D000565:Ameloblasts / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D003746:Dental Enamel Proteins / Q000276:immunology*; D015151:Immunoblotting; D008297:Male; D016253:Microscopy, Immunoelectron; D009804:Odontoblasts / Q000276:immunology; D051381:Rats; D017208:Rats, Wistar; D016610:Tissue Embedding / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D014746:Vimentin / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003746:Dental Enamel Proteins; D014746:Vimentin