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The Role of Receptor Tyrosine Kinases in Primordial Germ Cell Migration

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Stem Cell Migration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 750))

Abstract

During embryonic development in Drosophila, rodents, and other organisms, primordial germ cells (PGCs) migrate from their points of origin to the nascent gonads, where they give rise to germ line stem cells. Receptor tyrosine kinase (RTK) activity is required for normal migration of primordial germ cells in both Drosophila and rodents. In this chapter, we discuss in vivo as well as in vitro methods which have been used to elucidate the role of the RTK Torso in Drosophila germ cell migration. Included are protocols for embryo collection, fixation, and immunostaining; the dominant female sterile technique; in vitro culture and observation of PGCs; pole cell transplantation; and labeling of pole cells for in vivo observation.

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References

  1. Santos, A., and Lehmann, R. (2004) Germ cell specification and migration in Drosophila and beyond Current Biology 14, R578–R89.

    Google Scholar 

  2. Dadoune, J. (2007) New insights into male gametogenesis: what about the spermatogonial stem cell niche? Folia Histochemica et Cytobiologica 45a, 141–7.

    Google Scholar 

  3. Fuller, M., and Spradling, A. (2007) Male and female Drosophila germline stem cells: two versions of immortality Science 316, 402–4.

    Article  PubMed  CAS  Google Scholar 

  4. Li, J., Xia, F., and Li, W. (2003) Coactivation of STAT and Ras is required for germ cell proliferation and invasive migration in Drosophila Developmental Cell 5, 787–98.

    Article  PubMed  CAS  Google Scholar 

  5. Orth, J., Qui, J., Jester, W.J., and Pilder, S. (1997) Expression of the c-kit gene is critical for migration of neonatal rat gonocytes in vitro Biology of Reproduction 57, 676–83.

    Article  CAS  Google Scholar 

  6. Schlessinger, J. (2000) Cell signaling by receptor typrosine kinases Cell 103, 211–25.

    Article  PubMed  CAS  Google Scholar 

  7. Li, W. (2005) Functions and mechanisms of receptor tyrosine kinase Torso signaling: Lessons from Drosophila embryonic terminal development Developmental Dynamics 232, 656–72.

    Article  PubMed  CAS  Google Scholar 

  8. Brizzi, M., Dentelli, P,. Rosso, A., Yarden, Y., and Pegoraro, L. (1999) STAT protein recruitment and activation in c-Kit deletion mutants Journal of Biological Chemistry 274, 16965–72.

    Article  PubMed  CAS  Google Scholar 

  9. Deberry, C., Mou, S., and Linnekin, D. (1997) Stat1 associates with c-kit and is activated in response to stem cell factor Biochemical Journal 327, 73–80.

    PubMed  CAS  Google Scholar 

  10. Ning, Z., Li, J., McGuinness, M., and Arceci, R. (2001) STAT3 activation is required for Asp(816) mutant c-Kit induced tumorigenicity Oncogene 20, 4528–36.

    Article  PubMed  CAS  Google Scholar 

  11. Lu, X., Chou, T., Williams, N., Roberts, T., and Perrimon, N. (1993) Control of cell fate determination by p21ras/Ras1, an essential component of torso signaling in Drosophila Genes and Development 7, 621–32.

    Article  PubMed  CAS  Google Scholar 

  12. De Miguel, M., Cheng, L., Holland, E., Federspiel, M., and Donovan, P. (2002) Dissec-tion of the c-Kit signaling pathway in mouse primordial germ cells by retroviral-mediated gene transfer Proceedings of the National Academy of Sciences (USA) 99, 10458–63.

    Article  Google Scholar 

  13. Duffy, J., and Perrimon, N. (1994) The torso pathway in Drosophila: lessons on receptor tyrosine kinase signaling and pattern formation Developmental Biology 166, 380–95.

    Article  PubMed  CAS  Google Scholar 

  14. Perrimon, N., Lu, X., Hou, X., et al. (1995) Dissection of the Torso signal transduction pathway in Drosophila. Molecular Reproduction and Development 42, 515–22.

    Article  PubMed  CAS  Google Scholar 

  15. Starz-Gaiano, M., and Lehmann, R. (2001) Moving towards the next generation Mechanisms of Development 105, 5–18.

    Article  PubMed  CAS  Google Scholar 

  16. Wylie, C. (2000) Germ cells Current Opinion in Genetics and Development 10, 410–3.

    Article  PubMed  CAS  Google Scholar 

  17. Dahmann, C, ed. (2008) Drosophila Methods and Protocols. Totowa, NJ, Humana Press.

    Google Scholar 

  18. Sullivan, W., Ashburner, M., and Hawley, R. (2008) Drosophila Protocols. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory Press.

    Google Scholar 

  19. Sprenger, F., Stevens, L., and Nusslein-Volhard, C. (1989) The Drosophila gene torso encodes a putative receptor tyrorsine kinase Nature 338, 478–83.

    Article  PubMed  CAS  Google Scholar 

  20. Klingler, M., Erdelyi, M., Szabad, J., and Nusslein-Volhard, C. (1988) Function of torso in determining the terminal anlagen of the Drosophila embryo Nature 335, 275–7.

    Article  PubMed  CAS  Google Scholar 

  21. Hou, X., Chou, T., Melnick, M., and Perrimon, N. (1995) The torso receptor tyrosine kinase can activate Raf in a Ras-independent pathway Cell 81, 63–71.

    Article  PubMed  CAS  Google Scholar 

  22. Ambrosio, L., Mahowald, A., and Perrimon, N. (1989) Requirement of the Drosophila raf homologue for torso function Nature 342, 288–91.

    Article  PubMed  CAS  Google Scholar 

  23. Hou, X., Melnick, M., and Perrimon, N. (1996) Marelle acts downstream of the Drosophila HOP/JAK kinase and encodes a protein similar to the mammalian STATs Cell 84, 411–9.

    Article  PubMed  CAS  Google Scholar 

  24. Binari, R., and Perrimon, N. (1994) Stripe-specific regulation of pair-rule genes by hopscotch, a putative Jak family tyyrosine kinase in Drosophila Genes and Development 8, 300–12.

    Article  PubMed  CAS  Google Scholar 

  25. Harrison, D., McCoon, P., Binari, R., Gilman, M., and Perrimon, N. (1998) Drosophila unpaired enclodes a secreted protein that activates the JAK signaling pathway Genes and Development 12, 3252–63.

    Article  PubMed  CAS  Google Scholar 

  26. Harrison, D., Binari, R., Nahreini, T., Gilman, M., and Perrimon, N. (1995) Activation of a Drosophila Janus kinase (JAK) causes hematopoietic neoplasia and developmental defects EMBO Journal 14, 2857–65.

    PubMed  CAS  Google Scholar 

  27. Heimer, G., and Taylor, C. (1974) Improved mountant for immunofluorescence preparations Journal of Clinical Pathology 27, 254–6.

    Article  PubMed  CAS  Google Scholar 

  28. Chou, T., and Perrimon, N. (1992) Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila Genetics 151, 643–53.

    Google Scholar 

  29. Campos-Ortega, J., and Hartenstein, V. (1997) The Embryonic Development of Drosophila melanogaster. Berlin, Springer-Verlag.

    Google Scholar 

  30. Van Deusen, E. (1977) Sex determination in germ line chimeras of Drosophila melanogaster Journal of Embryology and Experimental Morphology 37, 173–85.

    PubMed  Google Scholar 

  31. Jaglarz, M., and Howard, K. (1994) Primordial germ cell migration in Drosophila melanogaster is controlled by somatic tissue Development 120, 83–9.

    PubMed  CAS  Google Scholar 

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Correspondence to Willis X. Li .

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Silver-Morse, L., Li, W.X. (2011). The Role of Receptor Tyrosine Kinases in Primordial Germ Cell Migration. In: Filippi, MD., Geiger, H. (eds) Stem Cell Migration. Methods in Molecular Biology, vol 750. Humana Press. https://doi.org/10.1007/978-1-61779-145-1_20

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  • DOI: https://doi.org/10.1007/978-1-61779-145-1_20

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-144-4

  • Online ISBN: 978-1-61779-145-1

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