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Gene Transfer to the Rodent Embryo by Retroviral Vectors

  • Grace K. Pavlath
  • Marla B. Luskin
Part of the Methods in Molecular Biology™ book series (MIMB, volume 97)

Abstract

Over the last 15 years investigators studying vertebrate development have capitalized on the use of retroviral-mediated gene transfer to determine the lineage relationships of diverse cell types, particularly in many regions of the mammalian central nervous system (for review, see ref. 1). Whereas an intraperitoneal or intrauterine injection of cell proliferation markers such as tritiated thymidine or bromodeoxyuridine suffices to examine the birthdates of cells, many fundamental questions dealing with the formation of a structure in the mammalian embryo often necessitate performing intrauterine surgery to introduce genes by retroviral vectors. As retroviral vectors can be used not only to study lineage (2), but also to introduce genes to perturb development (3, 4, 5), the methods for delivering retroviruses into the developing mammalian embryo will be in increasing demand. This chapter describes a set of procedures to generate and introduce retroviral vectors into rodent embryos.

Keywords

Chloral Hydrate Xiphoid Process Packaging Cell Line Acetate Sheet Biological Safety Cabinet 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    McDermott, K. W. and Luskin, M. B. (1995) The use of retroviral vectors in the study of cell lineage and migration during the development of the mammalian central nervous system, in Viral Vectors: Tools for Study and Genetic Manipulation of the Nervous System (Kaplitt, M. G. and Loewy, A. P., eds.), Academic, New York, pp. 411–433.Google Scholar
  2. 2.
    Sanes, J. R. (1989) Analysing cell lineage with a recombinant retrovirus. Trends Neurosci. 12, 21–28.PubMedCrossRefGoogle Scholar
  3. 3.
    Compere, S. J., Baldacci, P. A., Sharpe, A. H., and Jaenisch, R. (1989) Retroviral transduction of the human c-Ha-ras-1 oncogene into midgestation mouse embryos promotes rapid epithelial hyperplasia. Mol. Cell. Biol. 9, 6–14.PubMedGoogle Scholar
  4. 4.
    Galileo, D. S., Majors, J., Horwitz, A. F., and Sanes, J. R. (1992) Retrovirally introduced antisense integrin RNA inhibits neuroblast migration in vivo. Neuron 9, 1117–1131.PubMedCrossRefGoogle Scholar
  5. 5.
    Lillien, L. (1995) Changes in retinal cell fate induced by overexpression of EGF receptor. Nature 377, 158–162.PubMedCrossRefGoogle Scholar
  6. 6.
    Stoker, A. W. (1993) Retroviral vectors, in Retroviral Vectors (Davidson, A. J., and Elliot, R. M., eds.), Oxford Press, Oxford, UK, pp. 171–197.Google Scholar
  7. 7.
    Danos, O. and Mulligan, R. C. (1988) Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges. Proc. Natl. Acad. Sci. USA 85, 6460–6464.PubMedCrossRefGoogle Scholar
  8. 8.
    Pear, W. S., Nolan, G. P., Scott, M. L., and Baltimore, D. (1993) Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci. USA 90, 8392–8396.PubMedCrossRefGoogle Scholar
  9. 9.
    Sanes, J. R., Rubenstein, J. L., and Nicolas, J. F. (1986) Use of a recombinant retrovirus to study post implantation cell lineage in mouse embryos. EMBO J. 5, 3133–3142.PubMedGoogle Scholar
  10. 10.
    Price, J., Turner, D., and Cepko, C. (1987) Lineage analysis in the vertebrate nervous system by retrovirus-mediated gene transfer. Proc. Natl. Acad. Sci. USA 84, 156–160.PubMedCrossRefGoogle Scholar
  11. 11.
    Luskin, M. B., Parnavelas, J. G., and Barfield, J. A. (1993) Neurons, astrocytes, and oligodendrocytes of the rat cerebral cortex originate from separate progenitor cells: an ultrastructural analysis of clonally related cells. J. Neurosci. 13, 1730–1750.PubMedGoogle Scholar
  12. 12.
    Mione, M. C., Danevic, C., Boardman, P., Harris, B., and Parnavelas, J. G. (1994) Lineage analysis reveals neurotransmitter (GABA or glutamate) but not calcium-binding protein homogeneity in clonally related cortical neurons. J. Neurosci. 14, 107–123.PubMedGoogle Scholar
  13. 13.
    Luskin, M. B. (1993) Restricted proliferation and migration of postnatally generated neurons derived from the forebrain subventricular zone. Neuron 11, 173–189.PubMedCrossRefGoogle Scholar
  14. 14.
    Menezes, J. R. L., Smith, C. M., Nelson, K., and Luskin, M. B. (1995) The division of neuronal progenitor cells during migration in the neonatal mammalian forebrain. Mol. Cell. Neurosci. 6, 496–508.PubMedCrossRefGoogle Scholar
  15. 15.
    Turner, D. L. and Cepko, C. L. (1987) A common progenitor for neurons and glia persists in rat retina late in development. Nature 328, 131–136.PubMedCrossRefGoogle Scholar
  16. 16.
    Caggiano, M., Kauer, J. S., and Hunter, D. D. (1994) Globose basal cells are neuronal progenitors in the olfactory epithelium: a lineage analysis using a replication-incompetent retrovirus. Neuron 13, 339–352.PubMedCrossRefGoogle Scholar
  17. 17.
    Schwob, J. E., Huard, J. M., Luskin, M. B., and Youngentob, S. L. (1994) Retroviral lineage studies of the rat olfactory epithelium. Chem. Senses 19, 671–682.PubMedCrossRefGoogle Scholar
  18. 18.
    Bonnerot, C., Rocancourt, D., Briand, P., Grimber, G., and Nicolas, J. F. (1987) A beta-galactosidase hybrid protein targeted to nuclei as a marker for developmental studies. Proc. Natl. Acad. Sci. USA 84, 6795–6799.PubMedCrossRefGoogle Scholar
  19. 19.
    Kotani, H., Newton III, P. B., Zhang, S., Chiang, Y. L., Otto, E., Weaver, L., Blaese, R. M., Anderson, W. F., and McGarrity, G. J. (1994) Improved methods of retroviral vector transduction and production for gene therapy. Hum. Gene Ther. 5, 19–28.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  • Grace K. Pavlath
    • 1
  • Marla B. Luskin
    • 1
  1. 1.Department of Anatomy and Cell BiologyEmory University School of MedicineAtlanta

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