Glycoproteins in the retinal pigment epithelium of normal and dystrophic rats.
The apical membranes of retinal pigmented epithelium (RPE) were isolated from adult, normal (LE), and dystrophic (RCS) rats. The proteins of these RPE subfractions were separated through the use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lectin-binding properties of glycoproteins were examined in western blots through the use of lectin-peroxidase conjugates. No differences were detected between RPE membrane proteins from normal and dystrophic rats in silver-stained gels. However, these two preparations showed significant differences with respect to their binding of the lectins, Lens culinaris (Lentil), Tetragonolobus purpurea (Lotus), and concanavalin A (Con A). In particular, a glycoprotein with a molecular weight of 86 kD in the RPE apical membrane from normal rats bound Lentil, Lotus, and Con A, but in the membrane from dystrophic rats these binding sites were absent or significantly reduced. Another glycoprotein with a molecular weight of 175 kD was recognized by Lotus in the normal membrane preparation but not in the dystrophic RPE membrane preparation. Developmental studies show that these lectin-binding anomalies appear after postnatal day 11 and are, therefore, most likely coincident with eye opening in RCS rats. These results demonstrate that the RPE glycoproteins (86 and 175 kD) are significantly modified in dystrophic rats. The data also confirm previous observations that differences in the oligosaccharide chains, but not the polypeptide chains, of RPE membrane glycoproteins can be detected between normal and dystrophic rats. To the authors' knowledge, this is the first study to correlate developmentally regulated alterations in specific membrane-associated molecules in the RPE of dystrophic rats with the breakdown in phagocytosis that occurs in these rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Tien
L F
LF
Department of Anatomy and Neurobiology, College of Medicine, University of Tennessee, Memphis 38163.
McLaughlin
B J
BJ
Cooper
N G
NG
D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D007124:Immunoenzyme Techniques; D037102:Lectins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000378:metabolism; D008969:Molecular Sequence Data; D010857:Pigment Epithelium of Eye / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011922:Rats, Mutant Strains; D012162:Retinal Degeneration / Q000378:metabolism*
D037102:Lectins; D008562:Membrane Glycoproteins