Journal
DEVELOPMENTAL DYNAMICS
Volume 237, Issue 9, Pages 2575-2584Publisher
WILEY-LISS
DOI: 10.1002/dvdy.21683
Keywords
zebrafish; mutagenesis screen; lymphocyte; thymus; pharyngeal arch
Categories
Funding
- NHLBI [5 K08 HL04233-05]
- Huntsman Cancer Foundation [P30 CA042014]
- HHMI
- NIH [5 R01 HL48801-13]
- NATIONAL CANCER INSTITUTE [P30CA042014] Funding Source: NIH RePORTER
- NATIONAL CENTER FOR RESEARCH RESOURCES [R24RR014085] Funding Source: NIH RePORTER
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL048801, K08HL004233] Funding Source: NIH RePORTER
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Generation of mature T lymphocytes requires an intact hematopoietic stem cell compartment and functional thymic epithelium. We used the zebrafish (Danio rerio) to isolate mutations that affect the earliest steps in T lymphopoiesis and thymic organogenesis. Here we describe the results of a genetic screen in which gynogenetic diploid offspring from heterozygous females were analyzed by whole-mount in situ hybridization for the expression of rag-1. To assess immediately if a global defect in hematopoiesis resulted in the mutant phenotype, alpha-embryonic globin expression was simultaneously assayed for multilineage defects. In this report, we present the results obtained with this strategy and show representative mutant phenotypes affecting early steps in T-cell development and/or thymic epithelial cell development. We discuss the advantage of this strategy and the general usefulness of the zebrafish as a model system for vertebrate lymphopoiesis and thymic organogenesis.
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