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(Investigative Ophthalmology and Visual Science. 2004;45:4570-4575.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-0697
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Otx2 Homeobox Gene Induces Photoreceptor-Specific Phenotypes in Cells Derived from Adult Iris and Ciliary Tissue

Tadamichi Akagi,1,2 Michiko Mandai,1,3 Sotaro Ooto,1 Yasuhiko Hirami,1 Fumitaka Osakada,3,4 Ryoichiro Kageyama,2 Nagahisa Yoshimura,1 and Masayo Takahashi1,3

1From the Department of Ophthalmology and Visual Sciences, Graduate School of Medicine, the 2Institute for Virus Research, 3Department of Experimental Therapeutics, Translational Research Center, and 4Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan.

PURPOSE. It remains unclear which gene induction effectively generates photoreceptor-specific phenotypes from nonretinal tissues. The purpose of this study was to determine whether Crx and Otx2—homeobox genes related to photoreceptor development—can induce the generation of these phenotypes in cells derived from adult ciliary and iris tissue and in mesencephalon-derived neural stem cells.

METHODS. Crx and Otx2 were transferred into adult rat ciliary- and embryonic mesencephalon-derived neurospheres and adult rat iris-derived cells with the aid of a recombinant retrovirus. The presence of photoreceptor-specific phenotypes was confirmed by immunocytochemistry and Western blot analysis.

RESULTS. More than 90% of the Crx- and Otx2-transfected ciliary- and iris-derived cells exhibited rod opsin immunoreactivity, whereas few of the similarly transfected mesencephalon-derived neural stem cells expressed rod opsin. At least two additional key components of the phototransduction cascade, recoverin and G{partial}t1, were expressed by Crx- and Otx2-transfected iris-derived cells.

CONCLUSIONS. Crx and Otx2 effectively induced the generation of photoreceptor-specific phenotypes from ciliary- and iris-derived cells. That both Crx and Otx2 induced phenotype generation in cells derived from iris or ciliary tissue may suggest an approach to photoreceptor cell preparation for retinal transplantation.




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