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B.X. Wu, G. Gennadiy, Y. Chen, B. Rohrer, R.K. Crouch, J. Ma; Identification of RDH10, an All-trans Retinol Dehydrogenase, in Retinal Müller Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3517.
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Purpose: In vertebrates, 11-cis retinal is essential for maintaining normal visual function. Previous studies have shown that retinal Müller cells can synthesize 11-cis retinoids from all-trans retinol. The aim of this study is to investigate whether RDH10, an all-trans retinal dehydrogenase, previously identified only in the RPE (Wu et al., 2002), is expressed in retinal Müller cells. Methods: RT-PCR and immunoblot analysis were used to detect the RDH10 in rMC-1 cells, a rat Müller cell line. Retinol dehydrogenase activity assays were performed using the microsomal fraction of rMC-1 cells. The generation of all-trans retinal from tritiated all-trans retinol was measured by high pressure liquid chromatography. Immunohistochemistry was employed to analyze the cellular localization of RDH10 in mouse retinal sections, using a rabbit polyclonal antibody raised against the conserved C-terminal RDH10 peptide (position 327-341). Results: RDH10 was detected in rMC-1 cells by at both the RNA and protein levels by RT-PCR and immunoblot analysis. The identity of rat RDH10 was confirmed by sequence analysis of the RT-PCR product containing the full-length coding region isolated from the rMC-1 cells. The rat RDH10 cDNA sequence encodes a protein of 341 amino acids and shares 99.4 % homology with human, bovine and mouse RDH10 amino acid sequences. All-trans retinol dehydrogenase activity was also identified in the rMC-1 cells. The RDH activity prefers NADP as a co-factor, which is identical to the cofactor preference of the RDH10 in the RPE. Immunohistochemistry using RDH10 and CRALBP antibodies showed that RDH10 was colocalized with CRALBP in Müller cells in mouse retinal sections. Conclusions: RDH10 is expressed in retinal Müller cells in addition to the RPE. RDH10 generates all-trans retinal, which can serve as the substrate for the RGR photoisomerase in Müller cells.
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