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(Investigative Ophthalmology and Visual Science. 2004;45:3857-3862.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.03-1302
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Identification of RDH10, an All-trans Retinol Dehydrogenase, in Retinal Müller Cells

Bill X. Wu,1 Gennadiy Moiseyev,2 Ying Chen,1,2 Baerbel Rohrer,1,3 Rosalie K. Crouch,1 and Jian-xing Ma1,2

1From the Departments of Ophthalmology and 3Physiology and Neuroscience, Medical University of South Carolina, Charleston, South Carolina; and 2Department of Cell Biology, Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma.

PURPOSE. To investigate the expression of RDH10, an all-trans retinol dehydrogenase identified in the retinal pigment epithelium (RPE), in retinal Müller cells.

METHODS. The RDH10 protein levels in mouse eyecups and bovine tissues were examined by Western blot analysis using a polyclonal antibody against RDH10. The cellular localization in the retina was determined by immunohistochemistry. Expression of RDH10 in rMC-1, a cell line derived from rat Müller cells, was determined by RT-PCR and Western blot analysis. All-trans retinol dehydrogenase activity assays were performed using lysates from rMC-1 cells. The generation of all-trans retinal from tritiated all-trans retinol was analyzed by HPLC.

RESULTS. RDH10, retinal G protein-coupled receptor (RGR), and RPE65 all had higher expression levels in the eyecups of BALB/c than in C57Bl/6 mice. In addition to the RPE, RDH10 was also detected at lower levels in the retina and liver. Immunohistochemistry showed that RDH10 was localized in Müller cells in retinal sections. RDH10 was detected in rMC-1 cells, at both the RNA and protein levels. The rat RDH10 cDNA containing the full-length coding region was cloned from rMC-1 cells. The rat RDH10 cDNA encodes a protein of 341 amino acids and shares 99% sequence identity with human, bovine, and mouse RDH10 at the amino acid level. In rMC-1 cells, all-trans retinol dehydrogenase activity was detected in the microsomal fraction. NADP was shown to be the preferred cofactor, which is identical with the cofactor preference of the recombinant RDH10.

CONCLUSIONS. RDH10 was expressed in retinal Müller cells, in addition to the RPE. RDH10 generates all-trans retinal, which is the substrate for the photoisomerase RGR in Müller cells.




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