Neuron-specific enolase: a neuronal survival factor in the retinal extracellular matrix?
To identify soluble proteins of the retinal interphotoreceptor matrix (IPM), we isolated IPM from the bovine eye by gentle lavage and subjected it to SDS-PAGE. In the resultant gel, a 46 kDa band was particularly prominent and appeared to be a single protein. This protein was electroblotted to nitrocellulose membrane, digested with trypsin, and selected peptides were isolated by HPLC and subjected to Edman microsequencing. The amino acid sequences of the peptides were found to be virtually identical to that of human neuron-specific enolase (NSE). A monoclonal antibody specific for human NSE confirmed the presence of this enzyme in the bovine IPM by both Western blotting and immunocytochemical analysis. Immunofluorescence microscopy demonstrated that NSE is mainly localized to the basal domain of the IPM surrounding photoreceptor cells but is also prominent in the inner segments of the cone photoreceptor neurons. When NSE was added to cultures of human retinoblastoma cells, no effect on morphology was observed. However, a positive effect on cell growth and/or survival was readily apparent. It thus seems that not only is NSE a significant component of the retinal extracellular matrix, but that it could function as a survival (neuronotrophic) factor for photoreceptor neurons.
Li
A
A
Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892.
Lane
W S
WS
Johnson
L V
LV
Chader
G J
GJ
Tombran-Tink
J
J
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002454:Cell Differentiation; D002470:Cell Survival / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000378:metabolism*; D012160:Retina / Q000166:cytology / Q000502:physiology*
D000596:Amino Acids; D010751:Phosphopyruvate Hydratase