Effects of lethal irradiation in zebrafish and rescue by hematopoietic cell transplantation.
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 gamma 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 after 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.
Traver
David
D
Children's Hospital Boston and the Howard Hughes Medical Institute, 1 Blackfan Circle, Rm 07211, Boston, MA 02115, USA.
Winzeler
Alissa
A
Stern
Howard M
HM
Mayhall
Elizabeth A
EA
Langenau
David M
DM
Kutok
Jeffrey L
JL
Look
A Thomas
AT
Zon
Leonard I
LI
D000818:Animals; D030801:Animals, Genetically Modified; D005260:Female; D005720:Gamma Rays; D049452:Green Fluorescent Proteins; D006410:Hematopoiesis / Q000528:radiation effects*; D018380:Hematopoietic Stem Cell Transplantation / Q000401:mortality; D007668:Kidney / Q000166:cytology; D015458:Leukemia, T-Cell / Q000209:etiology / Q000473:pathology; D008164:Luminescent Proteins / Q000235:genetics; D008221:Lymphoid Tissue / Q000473:pathology / Q000528:radiation effects*; D008297:Male; D019172:Transplantation Conditioning; D014916:Whole-Body Irradiation; D015027:Zebrafish
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins