Detection of the Neu5 Ac (alpha 2,3) Gal (beta 1,4) GlcNAc sequence with the leukoagglutinin from Maackia amurensis: light and electron microscopic demonstration of differential tissue expression of terminal sialic acid in alpha 2,3- and alpha 2,6-linkage.
The Maackia amurensis leukoagglutinin has been shown to react specifically with the Neu5Ac (alpha 2,3) Gal sequence of asparagine-linked complex type oligosaccharides. We report here the preparation of Maackia amurensis lectin-gold complexes and their application for light and electron microscopic detection of the Neu5 Ac (alpha 2,3) Gal sequence in various tissues. The use of the lectin directly gold labeled was superior to a two-step cytochemical affinity technique using a fetuin-gold complex. The Maackia amurensis lectin-gold staining was inhibited by pre-incubation of the lectin-gold complexes with 50 mM alpha 2,3 sialyllactose, whereas alpha 2,6 sialyllactose up to concentrations of 1 M had no effect, thus demonstrating the high specificity of the histochemical staining. In addition to N-glycanase-sensitive asparagine-linked oligosaccharides, beta-elimination-sensitive serine/threonine-linked oligosaccharides could be detected. Data are presented which show that cellular staining patterns obtained with Maackia amurensis lectin-gold complexes may differ from those with elderberry bark lectin-gold, which detects the Neu5 Ac (alpha 2,6) Gal/GalN Ac sequence. Electron microscopic double labeling for direct study of the differential distribution of the Neu5 Ac (alpha 2,3) Gal and Neu5 Ac (alpha 2,6) Gal sequences is reported. Therefore, the availability of two sialic acid binding lectins with different linkage specificity for histochemistry provides the first opportunity to study tissue and cell type expression of these terminal sequences of glycoproteins.
Sata
T
T
Interdepartmental Electron Microscopy, University of Basal, Switzerland.
Lackie
P M
PM
Taatjes
D J
DJ
Peumans
W
W
Roth
J
J
D000117:Acetylglucosamine / Q000032:analysis / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002236:Carbohydrate Conformation; D002417:Cattle; D003106:Colon / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D005944:Glucosamine / Q000031:analogs & derivatives*; D006046:Gold / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods; D008854:Microscopy, Electron; D010179:Pancreas / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D010835:Phytohemagglutinins / Q000378:metabolism*; D037121:Plant Lectins; D051381:Rats; D012794:Sialic Acids / Q000032:analysis / Q000378:metabolism*; D013194:Staining and Labeling; D013363:Submandibular Gland / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D013552:Swine; D014197:Trees
D000596:Amino Acids; D010835:Phytohemagglutinins; D037121:Plant Lectins; D012794:Sialic Acids; C100156:leukoagglutinins, plants; D006046:Gold; D005944:Glucosamine; D000117:Acetylglucosamine