| Autor: |
Sheedlo, Harold J., Krishnamoorthy, Raghu, Nelson, Tammy, Agarwal, Neeraj, Liu, Jingjing, Roque, Rouel S., Wordinger, Robert J., Jaynes, David Jaynes C., Brun-Zinkelnagel, Anne-Marie, O'Brian, Paul, Aschenbrenner, John E., Turner, James E. |
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| Zdroj: |
Current Eye Research; Dec99, Vol. 19 Issue 6, 13 Black and White Photographs |
| Abstrakt: |
PURPOSE. This study was undertaken to determinethe pres-ence of retina-derived fetuin (RDF) protein and its message in retinaltissues and retinal pigment epithelial (RPE) cells. The techniques utilizedin this study included light micros-copy, immunochemistry, Western blot, reversetranscriptase-polymerase chain reaction (RT-PCR) and Southern blot. METHODS. Retinal tissues and sections from embryonic, early postnatal andadult normal rats and retinal pigment epithe-lial (RPE) cells from postnatalrats were immunostained for fetuin with a polyclonal fetuin antibody and aperoxidase conjugated-secondary antibody using immunocytochemistry and Westernblot analyses. The cDNA generated from RNA isolated from early postnatal ratretinas and RPE was probed with primers for rat fetuin, amplified by PCR andthe PCR products were analyzed by Southern blot. RESULTS. Fetuin (RDF) was immunolocalized to cells of the neuroepitheliumin retinas of early postnatal rats and most prominently in the nuclei andperinuclear region of cultured neonatal rat RPE cells. In adult retinas, ganglioncells, in-ner segments of photoreceptor cells, some components of the outerplexiform layer, ganglion cells and optic nerve pro-cesses were immunoreactivefor the fetuin protein. As shown by Western blot, fetuin (RDF) was higherin embryonic and early postnatal retinas than in late postnatal retinas, indicat-ingthat this protein may be developmentally regulated. Using RT-PCR, the messagefor rat fetuin was demonstrated in the retina and RPE of normal postnatalrats. Southern blot con-firmed that the PCR product from the retina and RPEwas generated from rat fetuin mRNA as well as from rat liver, the primarysource of fetuin. CONCLUSIONS. Fetuin, termed retina-derived fetuin (RDF), is reported forthe first time in retinal tissues. Fetuin is a cys-teine protease inhibitorthat may play a role in support of neuronal cell survival during early retinaldevelopment and the maintenance of neuronal activity. RDF ma... [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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