title: Ganglioside as an endogenous growth suppressor for glomerular mesangial cells.
abstract: BACKGROUND
  Glomerular mesangial cells potentially secrete many growth-modulating substances that could regulate mesangial cell proliferation. To date, however, the properties of such factors have not been fully evaluated.
  
  
  METHODS
  For that purpose, conditioned medium (CM) from mesangial cells was used for cross-feeding experiments. Cell proliferation was evaluated by 3H-thymidine incorporation assay and direct cell counting. The growth-regulatory molecule was further characterized using biochemical techniques.
  
  
  RESULTS
  Cross-feeding this CM to mesangial cells in vitro, despite stimulation with platelet-derived growth factor (PDGF), effectively suppressed the cells' synthesis of DNA in a dose-dependent manner. The inhibitory substance derived from mesangial cells was less than 3 kD in molecular mass, was heat stable, and was insensitive to proteinase K. After neuraminidase digestion, this inhibitory activity was lost. These data indicated that the inhibiting substance bore the typical features of gangliosides, which are multifunctional glycolipids that reside in cell membrane. Gangliosides were abundant in the CM from mesangial cells, as detected by metabolic radiolabeling and thin-layer chromatography (TLC). This result suggested that mesangial cells constitutively shed gangliosides. The growth suppressive activity in the CM was blunted when mesangial cells were pretreated with the ganglioside synthesis inhibitor d-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol-HCl (d-threo-PDMP; 20 micromol/L) in accordance with the decreased ganglioside content in cells. Finally, gangliosides isolated from CM of mesangial cells suppressed PDGF-induced DNA synthesis of mesangial cells.
  
  
  CONCLUSIONS
  These results suggest that mesangial cells constitutively shed gangliosides that then suppress the division of these cells in an autocrine-like manner.
authors.affiliation: Division of Kidney and Hypertension, Department of Internal Medicine, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo Japan. nobuotsuboi@aol.com
authors.forename: N
authors.initials: N
authors.lastname: Tsuboi
authors.forename: Y
authors.initials: Y
authors.lastname: Utsunomiya
authors.forename: T
authors.initials: T
authors.lastname: Kawamura
authors.forename: T
authors.initials: T
authors.lastname: Kawano
authors.forename: T
authors.initials: T
authors.lastname: Hosoya
authors.forename: T
authors.initials: T
authors.lastname: Ohno
authors.forename: H
authors.initials: H
authors.lastname: Yamada
mesh_terms: D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D005732:Gangliosides / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D005920:Glomerular Mesangium / Q000166:cytology* / Q000378:metabolism*; D006131:Growth Inhibitors / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D003470:Culture Media; D005732:Gangliosides; D006131:Growth Inhibitors