Histochemical detection of glycogen using Griffonia simplicifolia agglutinin II.
The utility of a lectin from Griffonia simplicifolia (GSA II) for demonstrating glycogen in situ was tested on fixed paraffin-embedded sections of a variety of tissues from rodents and man. The histochemical specificity of GSA II conjugated to horseradish peroxidase (GSA II-HRP) for glycogen was documented by the lability of such staining on adjacent sections treated with either malt diastase or alpha-amylase. In some tissue sites, the lectin-HRP conjugate imparted cytoplasmic staining that was diastase- and amylase-labile but resisted digestion with N-acetylglucosaminidase. Reactivity in the latter sites was attributed to glycogen and occurred in cell types having well-documented glycogen content, including liver hepatocytes, skeletal muscle fibres and polymorphonuclear leukocytes. In other tissue loci, GSA II binding was confined to cell surfaces or intracellular compartments, was not affected by prior diastase or amylase digestion, but was abolished with N-acetylglucosaminidase. Staining in these sites was attributed not to glycogen, but instead to glycoconjugate having sugar chains terminated with N-acetylglucosamine. These findings document the affinity of GSA II for glycogen in situ but do not conflict with the biochemically demonstrated affinity of GSA II for terminal N-acetylglucosamine. The results reported here show that the GSA II-HRP method may be useful for detecting physiological or pathological changes in the glycogen content of cells.
D000118:Acetylglucosaminidase / Q000378:metabolism; D000681:Amylases / Q000378:metabolism; D000818:Animals; D006003:Glycogen / Q000032:analysis*; D006651:Histocytochemistry; D006735:Horseradish Peroxidase; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008099:Liver / Q000032:analysis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D037121:Plant Lectins; D051381:Rats; D012756:Sheep
C045475:Griffonia simplicifolia lectins; D037102:Lectins; D037121:Plant Lectins; D006003:Glycogen; D006735:Horseradish Peroxidase; D000681:Amylases; D000118:Acetylglucosaminidase