title: Sertoli cell glycosylation patterns as affected by culture age and extracellular matrix.
abstract: This study evaluated the responsiveness of Sertoli cell glycosylation in vitro to changes in culture age and to the presence of a reconstituted basement membrane (Matrigel) or collagen IV/laminin substrata. Primary Sertoli cell cultures were prepared from 20-day-old rats and incubated with [3H]mannose, a monosaccharide specific for asparagine-linked oligosaccharides. The cells were harvested on Days 4, 6, or 10 of culture life. A supernatant enriched in cell-surface glycopeptides (the trypsinate) and a cell pellet stripped of surface glycoconjugates were evaluated separately. Glycopeptides derived from a Pronase digest of the two samples were fractionated using concanavalin-A lectin affinity chromatography into three major classes: multiantennary complex-type, biantennary complex-type, and high-mannose-type oligosaccharide structures. The proportion of radiolabeled glycopeptides appearing in each of the three classes did not differ between Days 4 and 6 of culture. In contrast, a significant increase in the percentage of radiolabeled glycopeptides containing multiantennary complex-type oligosaccharides was observed in cells harvested from the 10-day-old cultures. In other experiments, Sertoli cells were grown on various substrata: plastic; collagen IV/laminin; or Matrigel, a reconstituted basement membrane (RBM) composed of laminin, collagen IV, proteoglycan sulfate, entactin, and nidogen. Growth on RBM significantly increased multiantennary complex-type oligosaccharide formation compared to plastic, whereas the high-mannose-type glycopeptides increased in cells grown on collagen IV/laminin. These studies suggest that environmental and physiological conditions such as culture age and the presence of extracellular matrix significantly affect glycosylation patterns in Sertoli cell cultures.
authors: Department of Biology, Bucknell University, Lewisburg, Pennsylvania 17837.
authors: K C
authors: KC
authors: Page
authors: G J
authors: GJ
authors: Killian
authors: S E
authors: SE
authors: Nyquist
mesh_terms: D000375:Aging / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000502:physiology*; D006031:Glycosylation; D007797:Laminin / Q000494:pharmacology; D008297:Male; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000494:pharmacology; D010969:Plastics / Q000494:pharmacology; D011509:Proteoglycans / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology
chemical_list: D003470:Culture Media; D004338:Drug Combinations; D007797:Laminin; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010969:Plastics; D011509:Proteoglycans; C029967:nidogen; C059485:matrigel; D003094:Collagen