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Cell tracing shows the contribution of the yolk sac to adult haematopoiesis

Abstract

The first haematopoietic stem cells (HSCs) appear in the aorta-gonad-mesonephros (AGM) region, major vitelline and umbilical vessels, and placenta; however, whether they arise locally or from immigrant yolk sac precursor cells remains unclear. This issue is best addressed by measuring cell-lineage relationships rather than cell potentials. To undertake long-term in vivo tracing of yolk sac cells, we designed a non-invasive pulse-labelling system based on Cre/loxP recombination. Here we show that in Runx1+/- (runt-related transcription factor 1) heterozygous mice, yolk sac cells expressing Runx1 at embryonic day 7.5 develop into fetal lymphoid progenitors and adult HSCs. During mid-gestation the labelled (embryonic day 7.5) yolk sac cells colonize the umbilical cord, the AGM region and subsequently the embryonic liver. This raises the possibility that some HSCs associated with major embryonic vasculature are derived from yolk sac precursors. We observed virtually no contribution of the labelled cells towards the yolk sac vasculature, indicating early segregation of endothelial and haematopoietic lineages.

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Figure 1: Embryonic expression of Runx1 and analysis of inducible Runx1 -dependent cell tagging.
Figure 2: Embryo cell tagging kinetics at E7.5.
Figure 3: The embryonic haematopoietic progeny of yolk sac blood-island cells.
Figure 4: Representative β-gal staining of E10.5 dorsal aorta and E11.5 umbilical artery and vein sections of embryos activated at E7.5 by a single injection of 4′OHT.
Figure 5: The haematopoietic progeny in adult mice of yolk sac cells from the blood-island region.

References

  1. Medvinsky, A. & Dzierzak, E. Definitive hematopoiesis is autonomously initiated by the AGM region. Cell 86, 897–906 (1996)

    Article  CAS  Google Scholar 

  2. Gekas, C., Dieterlen-Lievre, F., Orkin, S. H. & Mikolla, H. K. A. The placenta is a niche for hematopoietic stem cells. Dev. Cell 8, 365–375 (2005)

    Article  CAS  Google Scholar 

  3. Cumano, A., Dieterlen-Lievre, F. & Godin, I. Lymphoid potential, probed before circulation in mouse, is restricted to caudal intraembryonic splanchnopleura. Cell 86, 907–916 (1996)

    Article  CAS  Google Scholar 

  4. Cumano, A., Ferraz, J. C., Klaine, M., Di Santo, J. P. & Godin, I. Intraembryonic, but not yolk sac hematopoietic precursors, isolated before circulation, provide long-term multilineage reconstitution. Immunity 15, 477–485 (2001)

    Article  CAS  Google Scholar 

  5. Yoder, M. C. et al. Characterization of definitive lymphohematopoietic stem cells in the day 9 murine yolk sac. Immunity 7, 335–344 (1997)

    Article  CAS  Google Scholar 

  6. Weissman, I., Papaioannou, V. & Gardner, R. Fetal Hematopoietic Origins of the Adult Hematolymphoid System (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1978)

    Google Scholar 

  7. Zhang, Y. et al. Inducible site-directed recombination in mouse embryonic stem cells. Nucleic Acids Res. 24, 543–548 (1996)

    Article  CAS  Google Scholar 

  8. Okuda, T., Van Deursen, J., Hiebert, S. W., Grosveld, G. & Downing, J. R. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell 84, 321–330 (1996)

    Article  CAS  Google Scholar 

  9. Wang, Q. et al. Disruption of the Cbfa2 gene causes necrosis and hemorrhaging in the central nervous system and blocks definitive hematopoiesis. Proc. Natl Acad. Sci. USA 93, 3444–3449 (1996)

    Article  CAS  ADS  Google Scholar 

  10. North, T. et al. Cbfa2 is required for the formation of intra-aortic hematopoietic clusters. Development 126, 2563–2575 (1999)

    CAS  PubMed  Google Scholar 

  11. Cai, Z. et al. Haploinsufficiency of AML1 affects the temporal and spatial generation of hematopoietic stem cells in the mouse embryo. Immunity 13, 423–431 (2000)

    Article  CAS  Google Scholar 

  12. Samokhvalov, I. M. et al. Multifunctional reversible knockout/reporter system enabling fully functional reconstitution of the AML1/Runx1 locus and rescue of hematopoiesis. Genesis 44, 115–121 (2006)

    Article  CAS  Google Scholar 

  13. Palis, J., Robertson, S., Kennedy, M., Wall, C. & Keller, G. Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. Development 126, 5073–5083 (1999)

    CAS  PubMed  Google Scholar 

  14. Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)

    Article  CAS  Google Scholar 

  15. Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001)

    Article  CAS  Google Scholar 

  16. Igarashi, H., Kouro, T., Yokota, T., Comp, P. C. & Kincade, P. W. Age and stage dependency of estrogen receptor expression by lymphocyte precursors. Proc. Natl Acad. Sci. USA 98, 15131–15136 (2001)

    Article  CAS  ADS  Google Scholar 

  17. Kellendonk, C. et al. Inducible site-specific recombination in the brain. J. Mol. Biol. 285, 175–182 (1999)

    Article  CAS  Google Scholar 

  18. Brotherton, T. W., Chui, D. H. K., Gauldie, J. & Patterson, M. Hemoglobin ontogeny during normal mouse fetal development. Proc. Natl Acad. Sci. USA 76, 2853–2857 (1979)

    Article  CAS  ADS  Google Scholar 

  19. Steiner, R. & Vogel, H. On the kinetics of erythroid cell differentiation in fetal mice: I. Microspectrophotometric determination of the hemoglobin content in erythroid cells during gestation. J. Cell. Physiol. 81, 323–338 (1973)

    Article  CAS  Google Scholar 

  20. Lien, E. A., Solheim, E. & Ueland, P. M. Distribution of tamoxifen and its metabolites in rat and human tissues during steady-state treatment. Cancer Res. 51, 4837–4844 (1991)

    CAS  PubMed  Google Scholar 

  21. Kisanga, E. R., Gjerde, J., Schjott, J., Mellgren, G. & Lien, E. A. Tamoxifen administration and metabolism in nude mice and nude rats. J. Steroid Biochem. Mol. Biol. 84, 361–367 (2003)

    Article  CAS  Google Scholar 

  22. Downs, K. M. & Davies, T. Staging of gastrulating mouse embryos by morphological landmarks in the dissecting microscope. Development 118, 1255–1266 (1993)

    CAS  Google Scholar 

  23. Nishikawa, S.-I. et al. In vitro generation of lymphohematopoietic cells from endothelial cells purified from murine embryos. Immunity 8, 761–769 (1998)

    Article  CAS  Google Scholar 

  24. Mao, X., Fujiwara, Y. & Orkin, S. H. Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice. Proc. Natl Acad. Sci. USA 96, 5037–5042 (1999)

    Article  CAS  ADS  Google Scholar 

  25. North, T. E. et al. Runx1 expression marks long-term repopulating hematopoietic stem cells in the midgestation mouse embryo. Immunity 16, 661–672 (2002)

    Article  CAS  Google Scholar 

  26. Minot, C. S. Development of the blood, the vascular system and the spleen. In Manual of Human Embryology (eds Keibel, F. and Mall, F. P.) 498–534 (J. B. Lippincott, Philadelphia, 1912)

    Google Scholar 

  27. Gerlai, R. Gene-targeting studies of mammalian behavior: is it the mutation or the background genotype? Trends Neurosci. 19, 177–181 (1996)

    Article  CAS  Google Scholar 

  28. Müller-Sieburg, C. E. & Riblet, R. Genetic control of the frequency of hematopoietic stem cells in mice: mapping of a candidate locus to chromosome 1. J. Exp. Med. 183, 1141–1150 (1996)

    Article  Google Scholar 

  29. Young, H. A. et al. Bone marrow and thymus expression of interferon-γ results in severe B-cell lineage reduction, T-cell lineage alterations, and hematopoietic progenitor deficiencies. Blood 89, 583–595 (1997)

    CAS  PubMed  Google Scholar 

  30. Yu, J.-M. et al. Expression of interferon-γ by stromal cells inhibits murine long-term repopulating hematopoietic stem cell activity. Exp. Hematol. 27, 895–903 (1999)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank S.-i. Aisawa for providing us with TT2 ES cells; F. Costantini for R26R-eYFP mice; N. Kazuki and J. Ure for their help with generating the knock-in mouse strains; and F. Melchers, A. Cumano and members of RIKEN CDB Kobe for critical discussion. I.M.S. was a recipient of a postdoctoral fellowship for foreign researchers from the Japan Society for the Promotion of Science. This work was supported in part by a grant for the Project for Realization of Regenerative Medicine (to S.-i.N.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Correspondence to Igor M. Samokhvalov.

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Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1–5 with Legends and detailed Supplementary Methods. The Supplementary Figures present additional data on the modified alleles used in the study as well as the scheme for Mer-Cre-Mer targeting into Runx1 locus. The Supplementary Figures also show the Runx1 expression in E7.5-E8.25 mouse concepti, the highly Runx1-positive cell clusters in day 8 - day 9 yolk sacs and provide the additional information on the analysis of the unspecific cell labeling. Supplementary Figure 5 shows the whole-mount X-Gal staining performed 12 hours after the 4’OHT administration at the nominal stage E8.0. The Supplementary Methods provide details about the Cre knock-in construct and generation of chimeric animals, induction with 4-hydroxitamoxifen, flow cytometry and cell preparations, whole-mount X-Gal staining and cryosectioning and PCR genotyping including the sequences of all primers used in this work. (PDF 4597 kb)

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Samokhvalov, I., Samokhvalova, N. & Nishikawa, Si. Cell tracing shows the contribution of the yolk sac to adult haematopoiesis. Nature 446, 1056–1061 (2007). https://doi.org/10.1038/nature05725

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