A retinal ganglion cell neurotrophic factor purified from the superior colliculus.
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.
Neurobiology Research Center, University of Sydney, New South Wales, Australia.
M
M
Schulz
T
T
Raju
G
G
Ralston
M R
MR
Bennett
D000818:Animals; D000831:Animals, Newborn; D001681:Biological Assay; D002470:Cell Survival; D002478:Cells, Cultured; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D011508:Chondroitin Sulfate Proteoglycans / Q000302:isolation & purification; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000302:isolation & purification; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000302:isolation & purification* / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D012160:Retina / Q000502:physiology*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000502:physiology*; D013477:Superior Colliculi / Q000032:analysis*
D011508:Chondroitin Sulfate Proteoglycans; D006023:Glycoproteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins