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(Investigative Ophthalmology and Visual Science. 2005;46:3411-3419.)
© 2005 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-1112
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Iris-Derived Cells from Adult Rodents and Primates Adopt Photoreceptor-Specific Phenotypes

Tadamichi Akagi,1,2 Joe Akita,1 Masatoshi Haruta,1 Takuya Suzuki,1 Yoshihito Honda,1 Tomoyuki Inoue,2 Shigeki Yoshiura,2 Ryoichiro Kageyama,2 Takaaki Yatsu,3 Masahiro Yamada,3 and Masayo Takahashi1,4

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

PURPOSE. The purpose of this study was to investigate the effects of various genes related to photoreceptor development on rodent and primate iris cells and the potential of iris cells as donor cells for retinal transplantation.

METHODS. Adult rat and monkey iris tissue were cultured in serum-free medium containing basic fibroblast growth factor. Gene deliveries of Crx, Nrl, NeuroD and some combinations (Crx-Nrl, Crx-NeuroD) were performed with recombinant retrovirus. Immunocytochemistry, Western blot analysis, RT-PCR, and intracellular recording were used to examine the expression of photoreceptor-specific phenotypes in the iris-derived cells after gene transfer, . Coculture of the iris-derived cells with embryonic retinal explant was conducted, to investigate the potential integration of these cells in coculture conditions.

RESULTS. Misexpression of Crx induced adult rat iris cells to express several photoreceptor-specific antigens and transcripts, such as rhodopsin, recoverin, cGMP-gated channel, arrestin, interphotoreceptor retinal-binding protein, rhodopsin kinase, and NeuroD. In primates, a combination of Crx and NeuroD was needed to induce monkey iris–derived cells to adopt photoreceptor-specific phenotypes. Furthermore, the photoreceptor-like cells derived from both rat- and primate-iris tissues showed rod photoreceptor-specific electrophysiological response to light stimuli after Crx and Crx-NeuroD gene transfer, respectively. The results further showed that iris-derived cells integrated in the developing host retina in coculture conditions.

CONCLUSIONS. Adult iris-derived cultured cells of both rodents and primates expressed photoreceptor-specific phenotypes by inductions of transcription factors. These iris-derived photoreceptor-like cells have electrophysiological characteristics of rod photoreceptors. Furthermore, they can integrate in the developing retina under coculture conditions.




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