Different rhodopsin monoclonal antibodies reveal different binding patterns on developing and adult rat retina.
We used a battery of 10 monoclonal antibodies directed against different identified peptide sequences within the carboxyl, transmembrane loop, and amino terminal regions of rhodopsin to label retinas from early postnatal and adult rats. Intensity of label, age of initial appearance of staining, and distribution of label varied depending on the antibody. Most antibodies showed detectable labeling at postnatal day 1, and were eventually observed binding to the cell bodies and the inner and outer segments of the photoreceptors. One amino terminal and two carboxyl terminal antibodies, however, showed no detectable labeling until postnatal day 5 and were only transiently detectable in the cell body region. These patterns cannot be explained by accessibility of binding site, binding affinity, fixation artifact, or crossreactivity. The results indicate that physiological and experimental parameters can alter the apparent immunocytochemical localization of conformationally active molecules such as rhodopsin. The results also suggest that rhodopsin can undergo light-dependent conformational changes in several different compartments within rat retinal photoreceptors before the time of eye opening.
Hicks
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Laboratory of Neurobiology, Rockefeller University, New York, New York 10021.
Barnstable
C J
CJ
D000375:Aging / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000276:immunology; D001666:Binding Sites, Antibody; D008027:Light; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010786:Photoreceptor Cells / Q000032:analysis; D011487:Protein Conformation / Q000528:radiation effects; D011863:Radioimmunoassay; D051381:Rats; D012160:Retina / Q000032:analysis / Q000254:growth & development*; D012168:Retinal Pigments / Q000032:analysis*; D012243:Rhodopsin / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D010446:Peptide Fragments; D012168:Retinal Pigments; D012243:Rhodopsin