Affinity cytochemical differentiation of glycoconjugates of small intestinal absorptive cells using Pisum sativum and Lens culinaris lectins.
We studied the subcellular localization of glycoconjugates recognized by the garden pea and lentil lectins (Pisum sativum, PSA; Lens culinaris, LCA) in mature absorptive cells of duodenum and jejunum of fasted rats. PSA and LCA are mannose-, glucose-, and N-acetyl-glucosamine-recognizing lectins that bind with high affinity to fucosylated core regions of N-glycosidically linked glycans. The binding reactions were cytochemically demonstrated in a pre-embedment incubation system using peroxidase-labeled lectins. Both pea and lentil lectins bound with constituents of nuclear envelope and endoplasmic reticulum, cisternae of the Golgi apparatus, several Golgi-associated vesicles, lysosomes, and portions of the plasma membrane. PSA and LCA label was non-homogeneous in the endoplasmic reticulum; in the Golgi apparatus the reactions were most intense in the cis and medial cisternae of the stacks. For inhibition of the intense reactions apparent in the Golgi apparatus, in lysosomes, and at the plasma membrane, considerably higher concentrations of competitive sugars were necessary than for abolition of the endoplasmic reticulum label. This indicates that endoplasmic reticulum glycoconjugates bind at low affinities with pea and lentil lectins, and that high-affinity PSA/LCA-binding glycoconjugates, which may correspond to corefucosylated N-linked glycans, predominate in cis and medial Golgi cisternae, lysosomes, and at the plasma membrane.
Pavelka
M
M
Institute of Micromorphology and Electron Microscopy, Vienna, Austria.
Ellinger
A
A
D000117:Acetylglucosamine / Q000378:metabolism; D000345:Affinity Labels / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D005947:Glucose / Q000378:metabolism; D006001:Glycoconjugates / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D006651:Histocytochemistry / Q000379:methods; D002479:Inclusion Bodies / Q000378:metabolism / Q000648:ultrastructure; D007421:Intestine, Small / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D037102:Lectins / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism / Q000648:ultrastructure; D008358:Mannose / Q000378:metabolism; D008854:Microscopy, Electron; D009685:Nuclear Envelope / Q000378:metabolism / Q000648:ultrastructure; D037121:Plant Lectins; D011134:Polysaccharides / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains
D000345:Affinity Labels; D006001:Glycoconjugates; D037102:Lectins; D037121:Plant Lectins; D011134:Polysaccharides; C033619:lentil lectin; C036292:pea lectin; D005947:Glucose; D008358:Mannose; D000117:Acetylglucosamine