Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Cryptic boundaries in roof plate and choroid plexus identified by intersectional gene activation

Nature Genetics volume 35, pages 70–75 (2003)Cite this article

Abstract

The hindbrain roof plate and choroid plexus are essential organizing centers for inducing dorsal neuron fates and sustaining neuron function. To map the formation of these structures, we developed a broadly applicable, high resolution, recombinase-based method for mapping the fate of cells originating from coordinates defined by intersecting combinations of expressed genes. Using this method, we show that distinct regions of hindbrain roof plate originate from discrete subdomains of rhombencephalic neuroectoderm expressing Wnt1; that choroid plexus, a secretory epithelium important for patterning later-formed hindbrain structures and maintaining neuron function1, derives from the same embryonic primordium as the hindbrain roof plate; and that, unlike the floor plate2,3, these dorsal organizing centers develop in a patterned, segmental manner, built from lineage-restricted compartments. Our data suggest that the roof plate and choroid plexus may be formed of functional units that are capable of differentially organizing the generation of distinct neuronal cell types at different axial levels.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: A genetic strategy for high-resolution fate mapping and its application to hindbrain development.
Figure 2: PLAP expression from the targeted R26::FLAP knock-in locus requires both Flpe and Cre activity, whereas eGFPF expression requires only Flpe.
Figure 3: A set of mouse strains expressing Flpe or Cre subdivides the embryonic hindbrain into an orthogonal grid of positional and gene expression information.
Figure 4: Switching on the lineage tracer PLAP at unique coordinate positions in the developing hindbrain.
Figure 5: The dual responsive indicator can be used to map development of hindbrain roof plate and fourth ventricle choroid plexus.

Similar content being viewed by others

References

  1. Yamamoto, M., McCaffery, P. & Drager, U.C. Influence of the choroid plexus on cerebellar development: analysis of retinoic acid synthesis. Brain Res. Dev. Brain Res. 93, 182–190 (1996).

    Article  CAS  Google Scholar 

  2. Fraser, S., Keynes, R. & Lumsden, A. Segmentation in the chick embryo hindbrain is defined by cell lineage restrictions. Nature 344, 431–435 (1990).

    Article  CAS  Google Scholar 

  3. Lumsden, A. & Keynes, R. Segmental patterns of neuronal development in the chick hindbrain. Nature 337, 424–428 (1989).

    Article  CAS  Google Scholar 

  4. Manzanares, M., Trainor, P.A., Ariza-McNaughton, L., Nonchev, S. & Krumlauf, R. Dorsal patterning defects in the hindbrain, roof plate and skeleton in the dreher (drJ) mouse mutant. Mech. Dev. 94, 147–156 (2000).

    Article  CAS  Google Scholar 

  5. Millonig, J.H., Millen, K.J. & Hatten, M.E. The mouse Dreher gene Lmx1a controls formation of the roof plate in the vertebrate CNS. Nature 403, 764–769 (2000).

    Article  CAS  Google Scholar 

  6. Lee, K.J., Dietrich, P. & Jessell, T.M. Genetic ablation reveals that the roof plate is essential for dorsal interneuron specification. Nature 403, 734–740 (2000).

    Article  CAS  Google Scholar 

  7. Zinyk, D., Mercer, E.H., Harris, E., Anderson, D.J. & Joyner, A.L. Fate mapping of the mouse midbrain-hindbrain constriction using a site-specific recombination system. Curr. Biol. 8, 665–668 (1998).

    Article  CAS  Google Scholar 

  8. Dymecki, S. & Tomasiewicz, H. Using Flp recombinase to characterize expansion of Wnt1-expressing neural progenitors in the mouse. Dev. Biol. 201, 57–65 (1998).

    Article  CAS  Google Scholar 

  9. Rodriguez, C.I. & Dymecki, S.M. Origin of the precerebellar system. Neuron 27, 475–486 (2000).

    Article  CAS  Google Scholar 

  10. Lumsden, A. & Krumlauf, R. Patterning the vertebrate neuraxis. Science 274, 1109–1115 (1996).

    Article  CAS  Google Scholar 

  11. Zambrowicz, B.P. et al. Disruption of overlapping transcripts in the ROSA βgeo 26 gene trap strain leads to widespread expression of β-galactosidase in mouse embryos and hematopoietic cells. Proc. Natl. Acad. Sci. USA 94, 3789–3794 (1997).

    Article  CAS  Google Scholar 

  12. Jiang, W. & Hunter, T. Analysis of cell-cycle profiles in transfected cells using a membrane-targeted GFP. Biotechniques 24, 349–354 (1998).

    Article  CAS  Google Scholar 

  13. Meyers, E.N., Lewandoski, M. & Martin, G.R. An Fgf8 mutant allelic series generated by Cre- and Flp-mediated recombination. Nat. Genet. 18, 136–141 (1998).

    Article  CAS  Google Scholar 

  14. Farley, F.W., Soriano, P., Steffen, L.S. & Dymecki, S.M. Widespread recombinase expression using FLPeR (flipper) mice. Genesis 28, 106–110 (2000).

    Article  CAS  Google Scholar 

  15. Bally-Cuif, L., Alvarado-Mallart, R.M., Darnell, D.K. & Wassef, M. Relationship between Wnt-1 and En-2 expression domains during early development of normal and ectopic met-mesencephalon. Development 115, 999–1009 (1992).

    CAS  PubMed  Google Scholar 

  16. Chai, Y. et al. Fate of the mammalian cranial neural crest during tooth and mandibular morphogenesis. Development 127, 1671–1679 (2000).

    CAS  PubMed  Google Scholar 

  17. Echelard, Y., Vassileva, G. & McMahon, A.P. Cis-acting regulatory sequences governing Wnt-1 expression in the developing mouse CNS. Development 120, 2213–2224 (1994).

    CAS  PubMed  Google Scholar 

  18. Frasch, M., Chen, X. & Lufkin, T. Evolutionary-conserved enhancers direct region-specific expression of the murine Hoxa-1 and Hoxa-2 loci in both mice and Drosophila. Development 121, 957–974 (1995).

    CAS  PubMed  Google Scholar 

  19. Soriano, P. Generalized lacZ expression with ROSA26 Cre reporter strain. Nat. Genet. 21, 70–71 (1999).

    Article  CAS  Google Scholar 

  20. Voiculescu, O., Charnay, P. & Schneider-Maunoury, S. Expression pattern of a Krox-20/Cre knock-in allele in the developing hindbrain, bones, and peripheral nervous system. Genesis 26, 123–126 (2000).

    Article  CAS  Google Scholar 

  21. Wilkinson, D.G., Bhatt, S., Chavrier, P., Bravo, R. & Charnay, P. Segment-specific expression of a zinc-finger gene in the developing nervous system of the mouse. Nature 337, 461–464 (1989).

    Article  CAS  Google Scholar 

  22. Prince, V. & Lumsden, A. Hoxa-2 expression in normal and transposed rhombomeres: independent regulation in the neural tube and neural crest. Development 120, 911–923 (1994).

    CAS  PubMed  Google Scholar 

  23. Altman, J. & Bayer, S. Development of the Cerebellar System: In relation to its evolution, structure, and functions (ed. Petralia, P.) (CRC Press, Boca Raton, 1997).

    Google Scholar 

  24. Wingate, R.J. & Hatten, M.E. The role of the rhombic lip in avian cerebellum development. Development 126, 4395–4404 (1999).

    CAS  PubMed  Google Scholar 

  25. Becker, N. et al. Several receptor tyrosine kinase genes of the Eph family are segmentally expressed in the developing hindbrain. Mech. Dev. 47, 3–17 (1994).

    Article  CAS  Google Scholar 

  26. Flenniken, A.M., Gale, N.W., Yancopoulos, G.D. & Wilkinson, D.G. Distinct and overlapping expression patterns of ligands for Eph-related receptor tyrosine kinases during mouse embryogenesis. Dev. Biol. 179, 382–401 (1996).

    Article  CAS  Google Scholar 

  27. Bergemann, A.D., Cheng, H.J., Brambilla, R., Klein, R. & Flanagan, J.G. ELF-2, a new member of the Eph ligand family, is segmentally expressed in mouse embryos in the region of the hindbrain and newly forming somites. Mol. Cell. Biol. 15, 4921–499 (1995).

    Article  CAS  Google Scholar 

  28. Inoue, T., Chisaka, O., Matsunami, H. & Takeichi, M. Cadherin-6 expression transiently delineates specific rhombomeres, other neural tube subdivisions, and neural crest subpopulations in mouse embryos. Dev. Biol. 183, 183–194 (1997).

    Article  CAS  Google Scholar 

  29. Gale, N.W. et al. Elk-L3, a novel transmembrane ligand for the Eph family of receptor tyrosine kinases, expressed in embryonic floor plate, roof plate and hindbrain segments. Oncogene 13, 1343–1352 (1996).

    CAS  PubMed  Google Scholar 

  30. Marin, F. & Puelles, L. Morphological fate of rhombomeres in quail/chick chimeras: a segmental analysis of hindbrain nuclei. Eur. J. Neurosci. 7, 1714–1738 (1995).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. Schneider-Maunoury for the Egr2::cre mouse strain; A. McMahon for plasmids pWEXP2 and pSP72MT34; T. Lufkin for the rhombomere 2–specific enhancer of Hoxa2; S. Thor for sequences encoding eGFPF; G. Martin for the pACTB::cre plasmid; J. Flanagan for plasmids encoding Eph receptors and their ligands; C. Tabin for a plasmid encoding cadherin-6; C. Tabin, C. Branda and A. Farago for critical reading of this manuscript; and S. Blackshaw for useful discussions. Work in the laboratories of S.M.D. and P.S. was supported by grants from the US National Institutes of Health. R.A. was supported by a Wills Foundation postdoctoral fellowship award and is currently supported by Taplin Funds for Discovery.

Author information

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Rajeshwar Awatramani, Carolyn Rodriguez, Jia Jia Mai & Susan M Dymecki

  2. Program in Developmental Biology and Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, 98109, Washington, USA

    Philippe Soriano

Authors
  1. Rajeshwar Awatramani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Philippe Soriano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carolyn Rodriguez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jia Jia Mai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Susan M Dymecki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Susan M Dymecki.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Awatramani, R., Soriano, P., Rodriguez, C. et al. Cryptic boundaries in roof plate and choroid plexus identified by intersectional gene activation. Nat Genet 35, 70–75 (2003). https://doi.org/10.1038/ng1228

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng1228

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing