Skip to main content
SpringerLink
Log in

Embryonic Intra-Aortic Clusters Undergo Myeloid Differentiation Mediated by Mesonephros-Derived CSF1 in Mouse

  • Published:
Stem Cell Reviews and Reports Aims and scope Submit manuscript

Abstract

The aorta-gonad-mesonephros (AGM) region contains intra-aortic clusters (IACs) thought to have acquired hematopoietic stem cell (HSC) potential in vertebrate embryos. To assess extrinsic regulation of IACs in the AGM region, we employed mouse embryos harboring a Sall1-GFP reporter gene, which allows identification of mesonephros cells based on GFP expression. Analysis of AGM region tissue sections confirmed mesonephros GFP expression. Mesonephric cells sorted at E10.5 expressed mRNA encoding Csf1, a hematopoietic cytokine, and corresponding protein, based on real-time PCR and immunocytochemistry, respectively. Further analysis indicated that some IACs express the CSF1 receptor, CSF1R. Expression of Cebpa and Irf8 mRNAs was higher in CSF1R-positive IACs, whereas that of Cebpε and Gfi1 mRNAs was lower relative to CSF1R-negative IACs, suggesting that CSF1/CSF1R signaling functions in IAC myeloid differentiation by modulating expression of these transcription factors. Colony formation assays using CSF1R-positive IACs revealed increased numbers of myeloid colonies in the presence of CSF1. Analysis using an intra-cellular signaling array indicated the greatest fold increase of Cleaved Caspase-3 in AGM cells in the presence of CSF1. Immunohistochemistry revealed that Cleaved Caspase-3 is primarily expressed in IACs in the AGM region, and incubation of IACs with CSF1 up-regulated Cleaved Caspase-3. Overall, our findings suggest that CSF1 secreted from mesonephros accelerates IAC myeloid differentiation in the AGM region, possibly via Caspase-3 cleavage.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Cumano, A., Dieterlen-Lievre, F., & Godin, I. (1996). Lymphoid potential, probed before circulation in mouse, is restricted to caudal intraembryonic splanchnopleura. Cell, 86(6), 907–916.

    Article  CAS  PubMed  Google Scholar 

  2. Ferkowicz, M. J., & Yoder, M. C. (2005). Blood island formation: longstanding observations and modern interpretations. Experimental Hematology, 33(9), 1041–1047.

    Article  PubMed  Google Scholar 

  3. Li, W., Johnson, S. A., Shelley, W. C., et al. (2003). Primary endothelial cells isolated from the yolk sac and Para-aortic splanchnopleura support the expansion of adult marrow stem cells in vitro. Blood, 102(13), 4345–4353.

    Article  CAS  PubMed  Google Scholar 

  4. McGrath, K. E., & Palis, J. (2005). Hematopoiesis in the yolk sac: more than meets the eye. Experimental Hematology, 33(9), 1021–1028.

    Article  PubMed  Google Scholar 

  5. Palis, J., Robertson, S., Kennedy, M., Wall, C., & Keller, G. (1999). Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. Development, 126(22), 5073–5084.

    CAS  PubMed  Google Scholar 

  6. Sugiyama, D., & Tsuji, K. (2006). Definitive hematopoiesis from endothelial cells in the mouse embryo; a simple guide. Trends in Cardiovascular Medicine, 16(2), 45–49.

    Article  CAS  PubMed  Google Scholar 

  7. Medvinsky, A., & Dzierzak, E. (1996). Definitive hematopoiesis is autonomously initiated by the AGM region. Cell, 86(6), 897–906.

    Article  CAS  PubMed  Google Scholar 

  8. Kumano, K., Chiba, S., Kunisato, A., et al. (2003). Notch1 but not Notch2 is essential for generating hematopoietic stem cells from endothelial cells. Immunity, 18(5), 699–711.

    Article  CAS  PubMed  Google Scholar 

  9. Yoder, M. C., Hiatt, K., Dutt, P., Mukherjee, P., Bodine, D. M., & Orlic, D. (1997). Characterization of definitive lymphohematopoietic stem cells in the day 9 murine yolk sac. Immunity, 7(3), 335–344.

    Article  CAS  PubMed  Google Scholar 

  10. Taoudi, S., Gonneau, C., Moore, K., et al. (2008). Extensive hematopoietic stem cell generation in the AGM region via maturation of VE-cadherin + CD45+ pre-definitive HSCs. Cell Stem Cell, 3(1), 99–108.

    Article  CAS  PubMed  Google Scholar 

  11. Rybtsov, S., Sobiesiak, M., Taoudi, S., et al. (2011). Hierarchical organization and early hematopoietic specification of the developing HSC lineage in the AGM region. The Journal of Experimental Medicine, 208(6), 1305–1315.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Sugiyama, D., Inoue-Yokoo, T., Fraser, S. T., Kulkeaw, K., Mizuochi, C., & Horio, Y. (2011). Embryonic regulation of the mouse hematopoietic niche. The Scientific World Journal, 11, 1770–1780.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Mizuochi, C., Fraser, S. T., Biasch, K., et al. (2012). Intra-aortic clusters undergo endothelial to hematopoietic phenotypic transition during early embryogenesis. PloS One, 7(4), e35763.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. North, T., Gu, T. L., Stacy, T., et al. (1999). Cbfa2 is required for the formation of intra-aortic hematopoietic clusters. Development, 126(11), 2563–2575.

    CAS  PubMed  Google Scholar 

  15. Cai, Z., de Bruijn, M., Ma, X., et al. (2000). Haploinsufficiency of AML1 affects the temporal and spatial generation of hematopoietic stem cells in the mouse embryo. Immunity, 13(4), 423–431.

    Article  CAS  PubMed  Google Scholar 

  16. Ling, K. W., Ottersbach, K., Van Hamburg, J. P., et al. (2004). GATA-2 plays two functionally distinct roles during the ontogeny of hematopoietic stem cells. The Journal of Experimental Medicine, 200(7), 871–882.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Lugus, J. J., Chung, Y. S., Mills, J. C., et al. (2007). GATA2 functions at multiple steps in hemangioblast development and differentiation. Development, 134(2), 393–405.

    Article  CAS  PubMed  Google Scholar 

  18. Bigas, A., Guiu, J., & Gama-Norton, L. (2013). Notch and Wnt signaling in the emergence of hematopoietic stem cells. Blood Cells, Molecules, and Diseases, 51(4), 264–270.

    Article  CAS  PubMed  Google Scholar 

  19. Desgrange, A., & Cereghini, S. (2015). Nephron patterning: lessons from Xenopus, zebrafish, and mouse studies. Cell, 4(3), 483–499.

    Article  Google Scholar 

  20. Takasato, M., Osafune, K., Matsumoto, Y., et al. (2004). Identification of kidney mesenchymal genes by a combination of microarray analysis and Sall1-GFP knockin mice. Mechanisms of Development, 121(6), 547–557.

    Article  CAS  PubMed  Google Scholar 

  21. Nishinakamura, R., & Takasato, M. (2005). Essential roles of Sall1 in kidney development. Kidney International, 68(5), 1948–1950.

    Article  CAS  PubMed  Google Scholar 

  22. Sasaki, T., Mizuochi, C., Horio, Y., et al. (2010). Regulation of hematopoietic cell clusters in the placental niche through SCF/kit signaling in embryonic mouse. Development, 137(23), 3941–3952.

    Article  CAS  PubMed  Google Scholar 

  23. McKinney-Freeman, S. L., Naveiras, O., Yates, F., et al. (2009). Surface antigen phenotypes of hematopoietic stem cells from embryos and murine embryonic stem cells. Blood, 114(2), 268–278.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Rosenbauer, F., & Tenen, D. G. (2007). Transcription factors in myeloid development: balancing differentiation with transformation. Nature Reviews Immunology, 7(2), 105–117.

    Article  CAS  PubMed  Google Scholar 

  25. Chittenden, T., Harrington, E. A., O'Connor, R., et al. (1995). Induction of apoptosis by the Bcl-2 homologue Bak. Nature, 374(6524), 733–736.

    Article  CAS  PubMed  Google Scholar 

  26. Oltval, Z. N., Milliman, C. L., & Korsmeyer, S. J. (1993). Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death. Cell, 74(4), 609–619.

    Article  Google Scholar 

  27. Bae, J., Leo, C. P., Hsu, S. Y., & Hsueh, A. J. (2000). MCL-1S, a splicing variant of the antiapoptotic BCL-2 family member MCL-1, encodes a proapoptotic protein possessing only the BH3 domain. Journal of Biological Chemistry, 275(33), 25255–25261.

    Article  CAS  PubMed  Google Scholar 

  28. Yang, J., Liu, X., Bhalla, K., et al. (1997). Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science, 275(5303), 1129–1132.

    Article  CAS  PubMed  Google Scholar 

  29. Robin, C., Ottersbach, K., Durand, C., et al. (2006). An unexpected role for IL-3 in the embryonic development of hematopoietic stem cells. Developmental Cell, 11(2), 171–180.

    Article  CAS  PubMed  Google Scholar 

  30. Petit-Cocault, L., Volle-Challier, C., Fleury, M., Péault, B., & Souyri, M. (2007). Dual role of Mpl receptor during the establishment of definitive hematopoiesis. Development, 134(16), 3031–3040.

    Article  CAS  PubMed  Google Scholar 

  31. Huang, X., Sakamoto, H., & Ogawa, M. (2009). Thrombopoietin controls proliferation of embryonic multipotent hematopoietic progenitors. Genes to Cells, 14(7), 851–860.

    Article  CAS  PubMed  Google Scholar 

  32. Crisan, M., Kartalaei, P. S., Vink, C., et al. (2015). BMP signalling differentially regulates distinct haematopoietic stem cell types. Nature Communications. doi:10.1038/ncomms9040.

    PubMed Central  Google Scholar 

  33. Robert-Moreno, À., Espinosa, L., de la Pompa, J. L., & Bigas, A. (2005). RBPjκ-dependent notch function regulates Gata2 and is essential for the formation of intra-embryonic hematopoietic cells. Development, 132(5), 1117–1126.

    Article  CAS  PubMed  Google Scholar 

  34. Robert-Moreno, À., Guiu, J., Ruiz-Herguido, C., et al. (2008). Impaired embryonic haematopoiesis yet normal arterial development in the absence of the notch ligand Jagged1. The EMBO Journal, 27(13), 1886–1895.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Durand, C., Robin, C., Bollerot, K., Baron, M. H., Ottersbach, K., & Dzierzak, E. (2007). Embryonic stromal clones reveal developmental regulators of definitive hematopoietic stem cells. Proceedings of the National Academy of Sciences, 104(52), 20838–20843.

    Article  CAS  Google Scholar 

  36. Peeters, M., Ottersbach, K., Bollerot, K., et al. (2009). Ventral embryonic tissues and hedgehog proteins induce early AGM hematopoietic stem cell development. Development, 136(15), 2613–2621.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Godin, I., Garcia-Porrero, J. A., Dieterlen-Lièvre, F., & Cumano, A. (1999). Stem cell emergence and hemopoietic activity are incompatible in mouse intraembryonic sites. The Journal of Experimental Medicine, 190(1), 43–52.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Guilbert, L. J., & Stanley, E. R. (1986). The interaction of 125I-colony-stimulating factor-1 with bone marrow-derived macrophages. Journal of Biological Chemistry, 261(9), 4024–4032.

    CAS  PubMed  Google Scholar 

  39. Yeung, Y. G., Jubinsky, P. T., Sengupta, A., Yeung, D., & Stanley, E. R. (1987). Purification of the colony-stimulating factor 1 receptor and demonstration of its tyrosine kinase activity. Proceedings of the National Academy of Sciences, 84(5), 1268–1271.

    Article  CAS  Google Scholar 

  40. Lin, H., Lee, E., Hestir, K., et al. (2008). Discovery of a cytokine and its receptor by functional screening of the extracellular proteome. Science, 320(5877), 807–811.

    Article  CAS  PubMed  Google Scholar 

  41. Mukouyama, Y. S., Hara, T., Xu, M. J., et al. (1998). In vitro expansion of murine multipotential hematopoietic progenitors from the embryonic aorta–gonad–mesonephros region. Immunity, 8(1), 105–114.

    Article  CAS  PubMed  Google Scholar 

  42. Dakic, A., Metcalf, D., Di Rago, L., Mifsud, S., Wu, L., & Nutt, S. L. (2005). PU. 1 regulates the commitment of adult hematopoietic progenitors and restricts granulopoiesis. The Journal of Experimental Medicine, 201(9), 1487–1502.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Scott, E. W., Fisher, R. C., Olson, M. C., Kehrli, E. W., Simon, M. C., & Singh, H. (1997). PU. 1 functions in a cell-autonomous manner to control the differentiation of multipotential lymphoid–myeloid progenitors. Immunity, 6(4), 437–447.

    Article  CAS  PubMed  Google Scholar 

  44. Iwasaki, H., Somoza, C., Shigematsu, H., et al. (2005). Distinctive and indispensable roles of PU. 1 in maintenance of hematopoietic stem cells and their differentiation. Blood, 106(5), 1590–1600.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Zhang, D. E., Zhang, P., Wang, N. D., Hetherington, C. J., Darlington, G. J., & Tenen, D. G. (1997). Absence of granulocyte colony-stimulating factor signaling and neutrophil development in CCAAT enhancer binding protein α-deficient mice. Proceedings of the National Academy of Sciences, 94(2), 569–574.

    Article  CAS  Google Scholar 

  46. Zhang, P., Iwasaki-Arai, J., Iwasaki, H., et al. (2004). Enhancement of hematopoietic stem cell repopulating capacity and self-renewal in the absence of the transcription factor C/EBPα. Immunity, 21(6), 853–863.

    Article  CAS  PubMed  Google Scholar 

  47. Yamanaka, R., Barlow, C., Lekstrom-Himes, J., et al. (1997). Impaired granulopoiesis, myelodysplasia, and early lethality in CCAAT/enhancer binding protein α-deficient mice. Proceedings of the National Academy of Sciences, 94(24), 13187–13192.

    Article  CAS  Google Scholar 

  48. Hock, H., Hamblen, M. J., Rooke, H. M., et al. (2003). Intrinsic requirement for zinc finger transcription factor Gfi-1 in neutrophil differentiation. Immunity, 18(1), 109–120.

    Article  CAS  PubMed  Google Scholar 

  49. Tamura, T., Nagamura-Inoue, T., Shmeltzer, Z., Kuwata, T., & Ozato, K. (2000). ICSBP directs bipotential myeloid progenitor cells to differentiate into mature macrophages. Immunity, 13(2), 155–165.

    Article  CAS  PubMed  Google Scholar 

  50. Scheller, M., Foerster, J., Heyworth, C. M., et al. (1999). Altered development and cytokine responses of myeloid progenitors in the absence of transcription factor, interferon consensus sequence binding protein. Blood, 94(11), 3764–3771.

    CAS  PubMed  Google Scholar 

  51. Richardson, E. T., Shukla, S., Nagy, N., Boom, W. H., Beck, R. C., Zhou, L., Landreth, G. E., Harding, C. V., (2015). ERK signaling is essential for macrophage development. PloS One, 10(10), e0140064.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Kelley, T. W., Graham, M. M., Doseff, A. I., Pomerantz, R. W., Lau, S. M., Ostrowski, M. C., Franke, T. F., Marsh, C. B., (1999). Macrophage colony-stimulating factor promotes cell survival through Akt/protein kinase B. Journal of Biological Chemistry, 274(37), 26393–26398.

    Article  CAS  PubMed  Google Scholar 

  53. Cohen, G. M. A. P. (1997). Caspases: the executioners of apoptosis. The Biochemical Journal, 326, 1–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Fujita, J., Crane, A. M., Souza, M. K., et al. (2008). Caspase activity mediates the differentiation of embryonic stem cells. Cell Stem Cell, 2(6), 595–601.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Janzen, V., Fleming, H. E., Riedt, T., et al. (2008). Hematopoietic stem cell responsiveness to exogenous signals is limited by caspase-3. Cell Stem Cell, 2(6), 584–594.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by a Grant-in-Aid for Exploratory Research by the Project for Realization of Regenerative Medicine, of the Ministry of Education, Culture, Sports, Science and Technology; and by a Bilateral grant to promote scientific exchange between Germany and Japan from the Japan Society for the Promotion of Science. We thank Dr. Elise Lamar for critical reading of our manuscript; Ms. Chiyo Yanagi-Mizuochi for technical support.

Author information

Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8577, Japan

    Tatsuya Sasaki, Junji Ishida & Akiyoshi Fukamizu

  2. Department of Research and Development of Next Generation Medicine, Faculty of Medical Sciences, Kyushu University, Fukuoka, 812-8582, Japan

    Yuka Tanaka, Kasem Kulkeaw, Ayako Yumine-Takai, Keai Sinn Tan & Daisuke Sugiyama

  3. Department of Cell Biology, Faculty of Medicine, Fukuoka University, Fukuoka, 814-0180, Japan

    Yuka Tanaka

  4. Department of Kidney Development, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto, 860-0811, Japan

    Ryuichi Nishinakamura

  5. Life Science Center, Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba, Ibaraki, 305-8577, Japan

    Junji Ishida & Akiyoshi Fukamizu

  6. Center for Clinical and Translational Research, Kyushu University, Fukuoka, 812-84582, Japan

    Daisuke Sugiyama

  7. Department of Clinical Study, Center for Advanced Medical Innovation, Kyushu University, Station for Collaborative Research 1 4F, 3-1-1 Maidashi, Higashi-Ku, Fukuoka, 812-8582, Japan

    Daisuke Sugiyama

Authors
  1. Tatsuya Sasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuka Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kasem Kulkeaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ayako Yumine-Takai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Keai Sinn Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ryuichi Nishinakamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Junji Ishida
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Akiyoshi Fukamizu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Daisuke Sugiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daisuke Sugiyama.

Ethics declarations

Conflict of Interest

The authors declare no potential conflicts of interest.

Electronic Supplementary Material

ESM 1

(DOCX 1379 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sasaki, T., Tanaka, Y., Kulkeaw, K. et al. Embryonic Intra-Aortic Clusters Undergo Myeloid Differentiation Mediated by Mesonephros-Derived CSF1 in Mouse. Stem Cell Rev and Rep 12, 530–542 (2016). https://doi.org/10.1007/s12015-016-9668-2

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12015-016-9668-2

Keywords

Search

Navigation