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Younghwa Shin, Gennadiy P Moiseyev, Krysten M Farjo, Yusuke Takahashi, Jian-Xing (Jay) Ma; RPE-specific knock-out of Retinol Dehydrogenase 10 altered retinoid profile. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3621.
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© ARVO (1962-2015); The Authors (2016-present)
Retinol dehydrogenase 10 (RDH10) is an enzyme required to generate retinoic acid during the early embryonic development, and its deletion leads to embryonic lethality. Although RDH10 is widely expressed during the embryonic development, its expression is mainly limited to the RPE in adulthood. We have generated mice with conditional knock-out of RDH10 to investigate its potential function in the retinoid metabolism in adult eyes.
RPE-specific knock-out mice of RDH10 (Rdh10RPE-/-) were generated by crossing Rdh10flox/flox with Best-1-Cre. Enucleated eyes from Rdh10RPE-/- and Rdh10flox/flox were dissected to obtain the eye cup, containing the RPE, choroid and sclera. The eye cups were used to confirm excision of RDH10 by both western blot and immunohistochemistry. We performed HPLC to analyze the retinoid profile from the eye ball samples of fully dark-adapted animals. Furthermore, we have analyzed the visual function of Rdh10RPE-/- by electroretinography (ERG).
Western blot results showed clean excision of RDH10 from the eye cup samples. Immunofluorescent staining in retinal cross-sections also confirmed RPE-specific excision of RDH10. HPLC retinoid profiling assay revealed Rdh10RPE-/- have altered retinoid profile compared to Rdh10flox/flox or Best-1-Cre alone. On the other hand, scotopic and photopic ERG results did not seem to vary with statistical significance.
We have successfully generated mice with RPE-specific knock-out of RDH10, and have observed altered retinoid profile in Rdh10RPE-/-. This model will allow us to further investigate the role(s) of RDH10 in the retinoid metabolism in adult eyes.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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