Single-cell imaging of normal and malignant cell engraftment into optically clear prkdc-null SCID zebrafish
- Moore, J.C., Tang, Q., Yordán, N.T., Moore, F.E., Garcia, E.G., Lobbardi, R., Ramakrishnan, A., Marvin, D.L., Anselmo, A., Sadreyev, R.I., Langenau, D.M.
- The Journal of experimental medicine 213(12): 2575-2589 (Journal)
- Registered Authors
- Langenau, David, Moore, John, Tang, Qin
- MeSH Terms
- Base Sequence
- Clone Cells
- DNA-Activated Protein Kinase/deficiency*
- DNA-Activated Protein Kinase/metabolism
- Disease Models, Animal
- Gamma Rays
- Imaging, Three-Dimensional/methods*
- Immunocompromised Host/radiation effects
- Luminescent Proteins/metabolism
- Muscle Cells/pathology
- Muscle Cells/radiation effects
- Neoplasm Transplantation*
- Optical Phenomena*
- Proto-Oncogene Proteins c-myc/metabolism
- Regeneration/radiation effects
- Single-Cell Analysis/methods*
- Transplantation, Homologous
- V(D)J Recombination/genetics
- Zebrafish Proteins/metabolism
- 27810924 Full text @ J. Exp. Med.
Moore, J.C., Tang, Q., Yordán, N.T., Moore, F.E., Garcia, E.G., Lobbardi, R., Ramakrishnan, A., Marvin, D.L., Anselmo, A., Sadreyev, R.I., Langenau, D.M. (2016) Single-cell imaging of normal and malignant cell engraftment into optically clear prkdc-null SCID zebrafish. The Journal of experimental medicine. 213(12):2575-2589.
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.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes