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Internalization of interphotoreceptor retinoid‐binding protein by the Xenopus retinal pigment epithelium
Author(s) -
Cunningham Lisa L.,
GonzalezFernandez Federico
Publication year - 2003
Publication title -
journal of comparative neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.855
H-Index - 209
eISSN - 1096-9861
pISSN - 0021-9967
DOI - 10.1002/cne.10861
Subject(s) - biology , retinal pigment epithelium , retina , microbiology and biotechnology , xenopus , visual phototransduction , retinal , immunoelectron microscopy , colocalization , anatomy , immunohistochemistry , biochemistry , neuroscience , immunology , gene
Xenopus rods and cones secrete into the interphotoreceptor matrix (IPM) a 124‐kDa glycoprotein termed interphotoreceptor retinoid‐binding protein (IRBP; Hessler et al. [1996] J. Comp. Neurol. 367:329–341). IRBP is confined to the IPM, being too large to diffuse through the zonulae adherentes between adjacent photoreceptor and Müller cells. Despite this physical entrapment within the subretinal space, IRBP is rapidly cleared from the IPM by an unknown mechanism. Immunohistochemistry and immunoelectron microscopy were used to localize IRBP in intact and detached retina‐retinal pigment epithelium (RPE) eyecups. The effects of light, dark, and time of day on the compartmentalization of IRBP were characterized by quantitative Western blot analysis and by immunoprecipitation of IRBP labeled in vivo by intraocular injection of [ 35 S]methionine. Immunohistochemistry showed that the apparent intercellular IRBP in both the RPE and the photoreceptors is resistant to saline extraction, in contrast to that in the IPM. In the RPE, IRBP was associated with matrix material within phagosomes and endosomes. The IPM, RPE, and retina contained 75%, 18%, and 7% of the total IRBP in the eye, respectively. The IPM and RPE contain 130 ± 14 pmoles and 34 ± 4 pmoles of IRBP, respectively. The amounts of IRBP in the RPE at middark and midlight were the same. Furthermore, the in vivo uptake of [ 35 S]methionine‐labeled IRBP was light independent. Our studies suggest that IRBP is not strictly confined to the subretinal space but rather that significant amounts are present intracellularly, particularly within the RPE, which does not synthesize IRBP. Furthermore, IRBP secreted by the photoreceptors is taken up from the IPM mainly through a light‐independent endocytic pathway separate from outer segment phagocytosis. The role of RPE endocytosis should be explored in relation to the function of IRBP. J. Comp. Neurol. 466:331–342, 2003. © 2003 Wiley‐Liss, Inc.