Oligosaccharide composition, localization, and developmental changes of a CNS-specific (F3-87-8) glycoprotein.
The F3-87-8 glycoprotein was isolated from rat brain by immunoaffinity chromatography after biosynthetic labeling by intracerebral administration of [3H]glucosamine, and the oligosaccharide composition of pronase-derived glycopeptides was determined by sequential lectin affinity chromatography and alkali treatment. Triantennary complex oligosaccharides (65%) and O-glycosidic oligosaccharides (18%) were the predominant types present, accompanied by 7-10% each of biantennary and high-mannose oligosaccharides. Twenty-two percent of the complex oligosaccharides had a fucose residue linked to the proximal N-acetylglucosamine of the chitobiose units. No poly(N-acetyllactosaminyl) or hybrid oligosaccharides were detected. Immunocytochemical studies on the localization of this glycoprotein in developing rat brain demonstrated that in 1-week postnatal cerebellum, there is light staining of the internal granule cell layer and surrounding the Purkinje cells. By 2 weeks, an intense staining of myelinating fiber tracts appears, accompanied by much lighter staining in the granule cell layer and at the base of the molecular layer. Staining of the white matter remains strong at 3 weeks postnatal, together with significant staining throughout the molecular layer, and then decreases in both areas by 1 month. In adult brain there is relatively uniform staining of approximately equal intensity in the white matter, granule cell layer, and molecular layer, whereas the Purkinje cell bodies appear unstained throughout development. In agreement with a previously reported immunochemical analysis, no staining was seen in other tissues, confirming the CNS-specific localization of this glycoprotein.
Department of Pharmacology, New York University Medical Center, New York 10016.
J A
JA
Ripellino
R U
RU
Margolis
R K
RK
Margolis
D000375:Aging; D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000254:growth & development* / Q000378:metabolism; D001923:Brain Chemistry; D002846:Chromatography, Affinity; D005944:Glucosamine / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification*; D006026:Glycoside Hydrolases; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins / Q000302:isolation & purification*; D009844:Oligosaccharides / Q000302:isolation & purification*; D009928:Organ Specificity; D051381:Rats
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D009419:Nerve Tissue Proteins; D009844:Oligosaccharides; D006026:Glycoside Hydrolases; D005944:Glucosamine