Expression patterns of the T antigen and the cryptic T antigen in rat fetuses: detection with the lectin amaranthin.
The lectin amaranthin, purified from the seeds of Amaranthus caudatus, has been shown to react specifically with the Gal beta 1,3GalNAc-alpha and the NeuAc alpha 2,3Gal beta 1,3GalNAc-alpha sequence which represent the T antigen and the cryptic T antigen, respectively. We report here the development of labeling techniques that apply amaranthin to stain paraffin sections from rat fetuses. Amaranthin staining was inhibited by pre-incubation of lectin-gold complexes with 10 mM Gal beta 1,3GalNAc-alpha-O-benzyl (synthetic T antigen) or 10 mM Gal beta 1,3GalNAc-alpha-O-aminophenylethyl-human serum albumin (T antigen neoglycoprotein), asialoglycophorin, asialofetuin, and asialomucin. The beta-elimination reaction also abolished the lectin staining demonstrating specificity for O-glycosidically linked structures. A comparison with monoclonal anti-T antigen antibody immunostaining demonstrated that amaranthin detects the T antigen and its cryptic form in tissue sections. Application of the galactose oxidase-Schiff sequence abolished amaranthin (and anti-T antibody) binding to the T antigen but not to its cryptic form, and therefore permitted their differentiation in tissue sections. Histochemical evidence was obtained indicating that amaranthin is a more specific anti-T reagent than peanut lectin. Data are presented that show the differential expression of the T antigen and the cryptic T antigen in organs and cells of rat fetuses late in gestation. Therefore, amaranthin can be used for histochemical detection of the T antigen and the cryptic T antigen, and facilitates discrimination between them.
Sata
T
T
Biocenter, University of Basel, Switzerland.
Zuber
C
C
Rinderle
S J
SJ
Goldstein
I J
IJ
Roth
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D001212:Asialoglycoproteins; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000032:analysis*; D005333:Fetus / Q000032:analysis* / Q000648:ultrastructure; D006046:Gold; D006651:Histocytochemistry; D007519:Isoantigens / Q000032:analysis*; D037102:Lectins; D008969:Molecular Sequence Data; D009928:Organ Specificity; D019887:Peanut Agglutinin; D037121:Plant Lectins; D051381:Rats; D054788:Ribosome Inactivating Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D014312:Trisaccharides / Q000032:analysis*
D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D001212:Asialoglycoproteins; D004187:Disaccharides; D007519:Isoantigens; D037102:Lectins; D019887:Peanut Agglutinin; D037121:Plant Lectins; D054789:Ribosome Inactivating Proteins, Type 1; C064068:Thomsen-Friedenreich antigen, cryptic; D014312:Trisaccharides; C061152:amaranthin protein, Amaranthus; C026503:asialoglycophorin; C016467:Thomsen-Friedenreich antigen; D006046:Gold; D054788:Ribosome Inactivating Proteins