Ultrastructural localization of mannoside residues on tissue sections: comparative evaluation of the enzyme-gold and the lectin-gold approaches.
Mannoside residues were revealed at the ultrastructural level in different cellular and extracellular compartments by means of the enzyme-gold and the lectin-gold approaches. For the enzyme-gold technique, an alpha-mannosidase-gold complex was prepared and conditions for the preparation of this complex as well as for its application were determined. Labeling was found over the rough endoplasmic reticulum mainly at the level of the membranes, the lumen of the cisternae being devoid of labeling. In the nucleus, the dense chromatin and the edge of the fibrillar threads in the nucleolus were intensely labeled. Few gold particles were present over the Golgi apparatus and mitochondria. The secretory granules in pancreatic cells, the peroxisomes in liver and the mucin in duodenal goblet cells were devoid of labeling. In the extracellular space, the basal lamina was labeled. Over the glomerular basal lamina, the labeling was mainly towards the epithelial side, in close contact with the podocytes. The results with the concanavalin A horseradish peroxidase (Con A-HRP)-gold technique were similar to those found with the enzyme-gold approach. Some differences were, however, detected at the level of the rough endoplasmic reticulum and the nucleus. In the endoplasmic reticulum, Con A-HRP-gold labeling was present over both the membranes and the lumen of the cisternae. In the nucleus, the labeling was mainly over the dispersed chromatin. These differences may be due to the binding of Con A not only to mannoside but also to other sugar residues as well as to the affinity of HRP-gold for some nucleoplasmic components.(ABSTRACT TRUNCATED AT 250 WORDS)
Londoño
I
I
Department of Anatomy, Faculty of Medicine, University of Montreal, Canada.
Bendayan
M
M
D000818:Animals; D003208:Concanavalin A; D004386:Duodenum / Q000032:analysis / Q000648:ultrastructure; D006027:Glycosides / Q000032:analysis*; D006046:Gold; D006651:Histocytochemistry; D007672:Kidney Cortex / Q000032:analysis / Q000648:ultrastructure; D008099:Liver / Q000032:analysis / Q000648:ultrastructure; D008361:Mannosidases; D008362:Mannosides / Q000032:analysis*; D010179:Pancreas / Q000032:analysis / Q000648:ultrastructure; D051381:Rats
D006027:Glycosides; D008362:Mannosides; D003208:Concanavalin A; D006046:Gold; D008361:Mannosidases