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Noninvasive Intravital Imaging of Thymocyte Dynamics in Medaka¹

Jie Li^2,*, Norimasa Iwanami^2,*, Vu Quynh Hoa^*, Makoto Furutani-Seiki and Yousuke Takahama^3,*

^* Division of Experimental Immunology, Institute for Genome Research, University of Tokushima, Tokushima, Japan; and Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, Kyoto, Japan

In vivo imaging of thymocytes has not been accomplished due to their localization deep within opaque body and high susceptibility to surgical stress. To overcome these problems, medaka is useful because of transparency and ex-uterine development. We report the noninvasive detection of thymocytes in transgenic medaka that express fluorescent protein under the control of immature-lymphocyte-specific rag1. We show that lymphoid progenitor cells colonize the thymus primordium in an anterior-to-posterior orientation-specific manner, revealing that extrathymic anterior components guide prevascular thymus colonization. We also show that developing thymocytes acquire "random walk motility" along with the expression of Ag receptors and coreceptors, suggesting that thymocyte walking is initiated at the developmental stage for repertoire selection. Thus, transgenic medaka enables real-time intravital imaging of thymocytes without surgical invasion.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This study was supported by grants-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology and Japan Society for the Promotion of Science Center-of-Excellence program (to N.I. and Y.T.).

² J.L. and N.I. contributed equally to this work.

³ Address correspondence and reprint requests to Dr. Yousuke Takahama, Division of Experimental Immunology, Institute for Genome Research, University of Tokushima, 770-8503 Tokushima, Japan. E-mail address: takahama{at}genome.tokushima-u.ac.jp

⁴ Abbreviations used in this paper: eGFP, enhanced GFP; wpf, weeks postfertilization; dpf, days postfertilization; UEA-1, Ulex europaeus agglutinin-1.

⁵ The online version of this article contains supplemental material.