Expression patterns of the T antigen and the cryptic T antigen in rat fetuses: detection with the lectin amaranthin

J Histochem Cytochem. 1990 Jun;38(6):763-74. doi: 10.1177/38.6.2335739.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Antigens, Tumor-Associated, Carbohydrate*
  • Asialoglycoproteins
  • Carbohydrate Sequence
  • Disaccharides / analysis*
  • Fetus / analysis*
  • Fetus / ultrastructure
  • Gold
  • Histocytochemistry
  • Isoantigens / analysis*
  • Lectins*
  • Molecular Sequence Data
  • Organ Specificity
  • Peanut Agglutinin
  • Plant Lectins*
  • Rats
  • Ribosome Inactivating Proteins
  • Ribosome Inactivating Proteins, Type 1
  • Trisaccharides / analysis*

Substances

  • Antibodies, Monoclonal
  • Antigens, Tumor-Associated, Carbohydrate
  • Asialoglycoproteins
  • Disaccharides
  • Isoantigens
  • Lectins
  • Peanut Agglutinin
  • Plant Lectins
  • Ribosome Inactivating Proteins, Type 1
  • Thomsen-Friedenreich antigen, cryptic
  • Trisaccharides
  • amaranthin protein, Amaranthus
  • asialoglycophorin
  • Thomsen-Friedenreich antigen
  • Gold
  • Ribosome Inactivating Proteins