Greatly impacting the field of neural development are new technologies for generating fate maps in mice and thus for illuminating relationships between embryonic and adult brain structures. Until now, efforts in mammalian models such as the mouse have presented challenges because their in utero development limits the access needed for traditional methods involving tracer injection or cell transplantation. But access is no longer an obstacle. It is now possible to deliver cell lineage tracers via noninvasive genetic, rather than physical, means. The hinge-pin of these new “genetic fate mapping” technologies is a class of molecule called a site-specific recombinase. The most commonly used being Cre and Flp. Through the capacity to produce precise DNA excisions, Cre or Flp can act as an on-switch, capable of transforming a silenced reporter transgene, for example, into a constitutively expressed one. A reporter transgene is, in effect, transformed by the excision event into an indelible cell-lineage tracer, marking ancestor and descendant cells. The actual cell population to be fate mapped is determined by recombinase parameters. Being genetically encoded, Cre or Flp is “delivered” to specific cells in the embryo using transgenics – promoter and enhancer elements from a gene whose expression is restricted to the desired cell type is used to drive recombinase expression. Thus, recombinase delivery is not only noninvasive but also restricted to specific embryonic cells based on their gene expression phenotype, lending molecular precision to the selection of cells for fate mapping. Resolution in cell type selection has recently been improved further by making lineage tracer activation dependent on two DNA excision events rather than just one. Here, in what is referred to as “intersectional genetic fate mapping,” lineage tracer is expressed only in those cells having undergone a Flp-dependent excision as well as a Cre-dependent excision, thus marking the embryonic cells lying at the intersection of two gene (Flp and Cre driver) expression domains. The field of hindbrain development, in particular, has seen great advances through application of these new approaches. For example, genetic fate maps of the cochlear nucleus have yielded surprising information about where in the embryonic hindbrain its constituent neurons arise and journey and what genes are expressed along the way. In this chapter, we detail materials and methods relevant to genetic fate mapping in general and intersectional genetic fate mapping in particular.
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References
Cepko, C. L., Austin, C. P., Walsh, C., Ryder, E. F., Halliday, A., and Fields- Berry, S. (1990) Studies of cortical development using retrovirus vectors. Cold Spring Harb. Symp. Quant. Biol. 55, 265–278.
Galileo, D. S., Gray, G. E., Owens, G. C., Majors, J., and Sanes, J. R. (1990) Neurons and glia arise from a common progenitor in chicken optic tectum: demonstration with two retroviruses and cell type-specific antibodies. Proc. Natl. Acad. Sci. USA 87, 458–462.
Walsh, C. and Cepko, C. L. (1988) Clonally related cortical cells show several migration patterns. Science 241, 1342–1345.
Wetts, R. F. and Fraser, S. E. (1988) Multipotent precursors can give rise to all major cell types of the frog retina. Science 239, 1142–1145.
Keller, R. E. (1975) Vital dye mapping of the gastrula and neuraula of Xenopus laevis. I. Prospective areas and morphogenetic movements of the superficial layer. Dev. Biol. 42, 222–241.
Le Douarin, N. (1982) The Neural Crest, Cambridge University Press, Cambridge.
Branda, C. and Dymecki, S. M. (2004) Talking about a revolution: The impact of site-specific recombinases on genetic analyses in mice. Develop. Cell 6, 7–28.
Joyner, A. and Zervas, M. (2006) Genetic inducible fate mapping in mouse: Establishing genetic lineages and defining genetic neuroanatomy in the nervous system. Dev. Dynamics 235, 2376–2385.
Dymecki, S. M. and Kim, J. C. (2007) Molecular neuroanatomy’s “Three Gs”: A Primer. Neuron 54, 17–34.
Dymecki, S. M. and Tomasiewicz, H. (1998) Using Flp-recombinase to characterize expansion of Wnt1-expressing neural progenitors in the mouse. Dev. Biol. 201, 57–65.
Zinyk, D., Mercer, E. H., Harris, E., Anderson, D. J., and Joyner, A. L. (1998) Fate mapping of the mouse midbrain-hindbrain constriction using a site-specific recombination system. Curr. Biol. 8, 665–668.
Rodriguez, C. I. and Dymecki, S. M. (2000) Origin of the precerebellar system. Neuron 27, 475–486.
Awatramani, R., Soriano, P., Rodriguez, C., Mai, J. J., and Dymecki, S. M. (2003) Cryptic boundaries in roof plate and choroid plexus identified by intersectional gene activation. Nat. Genet. 35, 70–75.
Farago, A., Awatramani, R., and Dymecki, S. (2006) Assembly of the brainstem cochlear complex is revealed by intersectional and subtractive genetic fate maps. Neuron 50,205–218.
Jensen, P., Farago, A. F., Awatramani, R. B., Scott, M. M., Deneris, E. S., Dymecki, S. M. (2008) Redefining the serotonergic system by genetic lineage. Nat Neurosci. April; 11(4), 417–419.
Wang, V. Y., Rose, M. F., and Zoghbi, H. Y. (2005) Math1 expression redefines the rhombic lip derivatives and reveals novel lineages within the brainstem and cerebellum. Neuron 48, 31–43.
Landsberg, R. L., Awatramani, R. B., Hunter, N. L., Farago, A. F., DiPietrantonio, H. J., Rodriguez, C. I., and Dymecki, S. M. (2005) Hindbrain rhombic lip is comprised of discrete progenitor cell populations allocated by Pax6. Neuron 48, 933–947.
Stark, W. M., Boocock, M. R., and Sherratt, D. J. (1992) Catalysis by site-specific recombinases. Trends Genet. 8, 432–439.
Dymecki, S. M. (2000) Site-specific recombination in cells and mice, in Gene Targeting: A Practical Approach (Joyner, A. L., ed.) second ed., Oxford University Press, Oxford,pp. 37–99.
Lumsden, A. and Krumlauf, R. (1996) Patterning the vertebrate neuraxis. Science 274, 1109–1115.
Rodriguez, C. I., Buchholz, F., Galloway, J., Sequerra, R., Kasper, J., Ayala, R., Stewart, A. F., and Dymecki, S. M. (2000) High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP. Nat. Genet. 25, 139–140.
Kosman, D., Mizutani, C. M., Lemons, D., Cox, W. G., McGinnis, W., and Bier, E. (2004) Multiplex detection of RNA expression in Drosophila embryos. Science 305, 846.
Tsien, J. Z., Chen, D. F., Gerber, D., Tom, C., Mercer, E. H., Anderson, D. J., Mayford, M., Kandel, E. R., and Tonegawa, S. (1996) Subregion- and cell type-restricted gene knockout in mouse brain. Cell 87, 1317–1326.
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Kim, J.C., Dymecki, S.M. (2009). Genetic Fate-Mapping Approaches: New Means to Explore the Embryonic Origins of the Cochlear Nucleus. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology™, vol 493. Humana Press. https://doi.org/10.1007/978-1-59745-523-7_5
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