Single-cell imaging of normal and malignant cell engraftment into optically clear prkdc-null SCID zebrafish
| Autor: | Qin Tang, John C. Moore, Nora Torres Yordán, Riadh Lobbardi, Elaine G. Garcia, Anthony Anselmo, Ruslan I. Sadreyev, Ashwin Ramakrishnan, Dieuwke L. Marvin, Finola E. Moore, David M. Langenau |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Optical Phenomena Immunology Cell DNA-Activated Protein Kinase Recombination-activating gene Proto-Oncogene Proteins c-myc Immunocompromised Host 03 medical and health sciences Imaging Three-Dimensional Live cell imaging medicine Animals Humans Regeneration Transplantation Homologous Immunology and Allergy Zebrafish Research Articles Muscle Cells Base Sequence biology Janus kinase 3 Homozygote Brief Definitive Report Anemia PRKDC Gene Zebrafish Proteins biology.organism_classification V(D)J Recombination Clone Cells 3. Good health Cell biology Transplantation Disease Models Animal Luminescent Proteins 030104 developmental biology medicine.anatomical_structure Gamma Rays Mutation Single-Cell Analysis Stem cell Neoplasm Transplantation |
| Zdroj: | The Journal of Experimental Medicine |
| ISSN: | 1540-9538 0022-1007 |
| DOI: | 10.1084/jem.20160378 |
| Popis: | Moore and colleagues present new strains of optically clear immune-deficient zebrafish that allow for dynamic imaging of regeneration and tumor progression at single-cell resolution in live animals. Cell transplantation into immunodeficient mice has revolutionized our understanding of regeneration, stem cell self-renewal, and cancer; yet models for direct imaging of engrafted cells has been limited. Here, we characterize zebrafish with mutations in recombination activating gene 2 (rag2), DNA-dependent protein kinase (prkdc), and janus kinase 3 (jak3). Histology, RNA sequencing, and single-cell transcriptional profiling of blood showed that rag2 hypomorphic mutant zebrafish lack T cells, whereas prkdc deficiency results in loss of mature T and B cells and jak3 in T and putative Natural Killer cells. Although all mutant lines engraft fluorescently labeled normal and malignant cells, only the prkdc mutant fish reproduced as homozygotes and also survived injury after cell transplantation. Engraftment into optically clear casper, prkdc-mutant zebrafish facilitated dynamic live cell imaging of muscle regeneration, repopulation of muscle stem cells within their endogenous niche, and muscle fiber fusion at single-cell resolution. Serial imaging approaches also uncovered stochasticity in fluorescently labeled leukemia regrowth after competitive cell transplantation into prkdc mutant fish, providing refined models to assess clonal dominance and progression in the zebrafish. Our experiments provide an optimized and facile transplantation model, the casper, prkdc mutant zebrafish, for efficient engraftment and direct visualization of fluorescently labeled normal and malignant cells at single-cell resolution. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|