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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Santos, A., and Lehmann, R. (2004) Germ cell specification and migration in Drosophila and beyond Current Biology 14, R578–R89.
Dadoune, J. (2007) New insights into male gametogenesis: what about the spermatogonial stem cell niche? Folia Histochemica et Cytobiologica 45a, 141–7.
Fuller, M., and Spradling, A. (2007) Male and female Drosophila germline stem cells: two versions of immortality Science 316, 402–4.
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.
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.
Schlessinger, J. (2000) Cell signaling by receptor typrosine kinases Cell 103, 211–25.
Li, W. (2005) Functions and mechanisms of receptor tyrosine kinase Torso signaling: Lessons from Drosophila embryonic terminal development Developmental Dynamics 232, 656–72.
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.
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.
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.
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.
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.
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.
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.
Starz-Gaiano, M., and Lehmann, R. (2001) Moving towards the next generation Mechanisms of Development 105, 5–18.
Wylie, C. (2000) Germ cells Current Opinion in Genetics and Development 10, 410–3.
Dahmann, C, ed. (2008) Drosophila Methods and Protocols. Totowa, NJ, Humana Press.
Sullivan, W., Ashburner, M., and Hawley, R. (2008) Drosophila Protocols. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory Press.
Sprenger, F., Stevens, L., and Nusslein-Volhard, C. (1989) The Drosophila gene torso encodes a putative receptor tyrorsine kinase Nature 338, 478–83.
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.
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.
Ambrosio, L., Mahowald, A., and Perrimon, N. (1989) Requirement of the Drosophila raf homologue for torso function Nature 342, 288–91.
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.
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.
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.
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.
Heimer, G., and Taylor, C. (1974) Improved mountant for immunofluorescence preparations Journal of Clinical Pathology 27, 254–6.
Chou, T., and Perrimon, N. (1992) Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila Genetics 151, 643–53.
Campos-Ortega, J., and Hartenstein, V. (1997) The Embryonic Development of Drosophila melanogaster. Berlin, Springer-Verlag.
Van Deusen, E. (1977) Sex determination in germ line chimeras of Drosophila melanogaster Journal of Embryology and Experimental Morphology 37, 173–85.
Jaglarz, M., and Howard, K. (1994) Primordial germ cell migration in Drosophila melanogaster is controlled by somatic tissue Development 120, 83–9.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-61779-145-1_20
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-144-4
Online ISBN: 978-1-61779-145-1
eBook Packages: Springer Protocols