Localization of cellular retinoic acid-binding protein to amacrine cells of rat retina.
Monoclonal antibodies to performic acid-oxidized cellular retinoic acid-binding protein (CRABP) from bovine retina were prepared by fusion of spleen cells from immunized mice with mouse myeloma cells. Five antibodies were studied in detail. It was established by ELISA that the antibodies react with CRABP and oxidized CRABP, but not with other oxidized or unmodified retinoid-binding proteins. Competitive ELISA demonstrated that the antibodies react with heat-denatured antigen but not with native protein. Western blotting and immunostaining, following sodium dodecyl sulfate gel electrophoresis, provided evidence for recognition of a single component in retinal supernatants whose staining is prevented by preabsorption of the antibody with heat-denatured CRABP. The insoluble fraction from a retinal homogenate contains residual CRABP and two weakly-reacting components, whose staining is not affected by preabsorption of the antibody with antigen. Each antibody produces the same staining pattern on cryostat sections of rat retina by indirect immunofluorescence. Amacrine somata on both sides of the inner plexiform layer are labeled, as well as processes forming laminae within this layer. These results suggest that retinoic acid may play a functional role in the inner retina.
Gaur
V P
VP
Department of Ophthalmology, School of Medicine, University of Washington, Seattle 98195.
de Leeuw
A M
AM
Milam
A H
AH
Saari
J C
JC
D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D009474:Neurons / Q000032:analysis*; D051381:Rats; D018168:Receptors, Retinoic Acid; D012160:Retina / Q000032:analysis*; D014212:Tretinoin
D002352:Carrier Proteins; D018168:Receptors, Retinoic Acid; D014212:Tretinoin