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Molecular and Cellular Biology, May 2004, p. 4513-4521, Vol. 24, No. 10
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.10.4513-4521.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Specification of the Retinal Fate of Mouse Embryonic Stem Cells by Ectopic Expression of Rx/rax, a Homeobox Gene

Yoko Tabata,1 Yasuo Ouchi,1 Haruyuki Kamiya,2 Toshiya Manabe,2,3 Ken-ichi Arai,1 and Sumiko Watanabe1*

Division of Molecular and Developmental Biology,1 Division of Neuronal Network, Department of Basic Medical Sciences, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639,3 Division of Cell Biology and Neurophysiology, Department of Neuroscience, Faculty of Medicine, Kobe University, Chuo-ku, Kobe 650-0017, Japan2

Received 26 November 2003/ Returned for modification 1 February 2004/ Accepted 11 February 2004

With the goal of generating retinal cells from mouse embryonic stem (ES) cells by exogenous gene transfer, we introduced the Rx/rax transcription factor, which is expressed in immature retinal cells, into feeder-free mouse ES cells (CCE). CCE cells expressing Rx/rax as well as enhanced green fluorescent protein (CCE-RX/E cells) proliferated and remained in the undifferentiated state in the presence of leukemia inhibitory factor, as did parental ES cells. We made use of mouse embryo retinal explant cultures to address the differentiation ability of grafted ES cells. Dissociated embryoid bodies were treated with retinoic acid for use as donor cells and cocultured with retina explants for 2 weeks. In contrast to the parental CCE cells, which could not migrate into host retinal cultures, CCE-RX/E cells migrated into the host retina and extended their process-like structures between the host retinal cells. Most of the grafted CCE-RX/E cells became located in the ganglion cell and inner plexiform layers and expressed ganglion and horizontal cell markers. Furthermore, these grafted cells had the electrophysiological properties expected of ganglion cells. Our data thus suggest that subpopulations of retinal neurons can be generated in retinal explant cultures from grafted mouse ES cells ectopically expressing the transcription factor Rx/rax.

* Corresponding author. Mailing address: Department of Molecular and Developmental Biology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Phone: 81-3-5449-5663. Fax: 81-3-5449-5474. E-mail: sumiko{at}ims.u-tokyo.ac.jp.

Molecular and Cellular Biology, May 2004, p. 4513-4521, Vol. 24, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.10.4513-4521.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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