A retinal ganglion cell neurotrophic factor purified from the superior colliculus

J Neurochem. 1990 Sep;55(3):832-41. doi: 10.1111/j.1471-4159.1990.tb04567.x.

Abstract

Dissociated neonatal rat retinal ganglion cells can be maintained by the addition of an extract from the neonatal superior colliculus. This extract can support 95% of ganglion cells over 24 h in culture; in addition it promotes the expression of neurites from these cells. This report describes the purification of a neurotrophic factor from the superior colliculus which supports the survival of 80% of retinal ganglion cells over 24 h in vitro. The purification procedure involves a combination of dye-ligand, anion-exchange, and molecular sieve chromatography. The purified neurotrophic factor has a Stokes radius of approximately 200 A using molecular sieve chromatography in the presence of a chaotropic agent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified factor indicates that it is a glycoprotein that migrates with a molecular mass greater than 400 kDa. Further characterization of this high-molecular-mass glycoprotein by enzymatic digestion demonstrated that it is a chondroitin sulfate proteoglycan. This factor is clearly distinguishable from other neurotrophic factors that have an effect on retinal ganglion cells such as brain-derived neurotrophic factor and fibroblast growth factor. The chondroitin sulfate proteoglycan from the neonatal superior colliculus is the first proteoglycan to be identified as a neurotrophic factor.

MeSH terms

  • Animals
  • Animals, Newborn
  • Biological Assay
  • Cell Survival
  • Cells, Cultured
  • Chemical Phenomena
  • Chemistry, Physical
  • Chondroitin Sulfate Proteoglycans / isolation & purification
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique
  • Glycoproteins / isolation & purification
  • Molecular Weight
  • Nerve Growth Factors / isolation & purification*
  • Nerve Growth Factors / pharmacology
  • Nerve Tissue Proteins / isolation & purification*
  • Nerve Tissue Proteins / pharmacology
  • Rats
  • Retina / physiology*
  • Retinal Ganglion Cells / cytology
  • Retinal Ganglion Cells / physiology*
  • Superior Colliculi / analysis*

Substances

  • Chondroitin Sulfate Proteoglycans
  • Glycoproteins
  • Nerve Growth Factors
  • Nerve Tissue Proteins