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Blood, 1 September 2004, Vol. 104, No. 5, pp. 1298-1305.
Prepublished online as a Blood First Edition Paper on May 13, 2004; DOI 10.1182/blood-2004-01-0100.
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Submitted January 13, 2004
Accepted April 22, 2004
Biological effects of lethal irradiation and rescue by hematopoietic cell transplantation in zebrafish
David Traver, Alissa Winzeler, Howard M Stern, Elizabeth A Mayhall, David M Langenau, Jeffrey L Kutok, A T Look, and Leonard I Zon*
Department of Medicine, Childrens Hospital Boston, Boston, MA, USA; Howard Hughes Medical Institute, Boston, MA, USA
Department of Medicine, Childrens Hospital Boston, Boston, MA, USA; Howard Hughes Medical Institute, Boston, MA, USA; Department of Pathology, Brigham and Womens Hospital, Boston, MA, USA
Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA
Department of Pathology, Brigham and Womens Hospital, Boston, MA, USA
* Corresponding author; email: zon{at}enders.tch.harvard.edu.
The study of hematopoiesis has been greatly facilitated by transplantation of blood cell populations into recipient animals. Efficient engraftment of donor cells generally requires ablation of the host hematopoietic system. The zebrafish has recently emerged as a developmental and genetic system to study hematopoiesis. To enable the study of hematopoietic stem cell (HSC) biology, immune cell function, and leukemogenesis in zebrafish, we have developed hematopoietic cell transplantation (HCT) into adult recipient animals conditioned by  irradiation. Dose-response experiments showed that the minimum lethal dose (MLD) of 40 Gy led to the specific ablation of hematolymphoid cells and death by 14 days post-irradiation. Sublethal irradiation doses of 20 Gy predominantly ablated lymphocytes and permitted transplantation of a lethal T cell leukemia. Finally, transplantation of hematopoietic cells carrying transgenes yielding red fluorescent erythrocytes and green fluorescent leukocytes showed that HCT is sufficient to rescue the MLD, that recipient hematolymphoid tissues were repopulated by donor-derived cells, and that donor blood cell lineages can be independently visualized in living recipients. Together, these results establish transplantation assays to test for HSC function and oncogenic transformation in zebrafish.
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