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.

A role for Gbx2 in repression of Otx2 and positioning the mid/hindbrain organizer

Abstract

The mid/hindbrain (MHB) junction can act as an organizer to direct the development of the midbrain and anterior hindbrain1,2. In mice, Otx2 is expressed in the forebrain and midbrain and Gbx2 is expressed in the anterior hindbrain, with a shared border at the level of the MHB organizer. Here we show that, in Gbx2-/- mutants, the earliest phenotype is a posterior expansion of the Otx2 domain during early somite stages. Furthermore, organizer genes are expressed at the shifted Otx2 border, but not in a normal spatial relationship. To test whether Gbx2 is sufficient to position the MHB organizer, we transiently expressed Gbx2 in the caudal Otx2 domain and found that the Otx2 caudal border was indeed shifted rostrally and a normal appearing organizer formed at this new Otx2 border. Transgenic embryos then showed an expanded hindbrain and a reduced midbrain at embryonic day 9.5–10. We propose that formation of a normal MHB organizer depends on a sharp Otx2 caudal border and that Gbx2 is required to position and sharpen this border.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1: Transformation of the metencephalon towards a mesencephalic fate at ESS in Gbx2 mutants.
Figure 2: Otx2 and Wnt1 are repressed and Gbx2 and Fgf8 are induced in the caudal midbrain at ESS in Wnt1–Gbx2 transgenics.
Figure 3: The sizes of the mesencephalon and metencephalon are altered transiently in Wnt1–Gbx2 transgenics.
Figure 4: Schematic representation of the Otx2, Gbx2, Wnt1 and Fgf8 domains of expression in wild-type, Gbx2 mutant and Wnt1–Gbx2 transgenic embryos at the 6-somite stage.

References

  1. Wassef,M. & Joyner,A. L. Early mesencephalon/metencephalon patterning and development of the cerebellum. Perspect. Dev. Neurobio. 5, 3–16 (1997).

    CAS  Google Scholar 

  2. Alvarado-Mallart,R. M. Fate and potentialities of the avian mesencephalic/metencephalic neuroepithelium. J. Neurobiol. 24, 1341–1355 (1993).

    Article  CAS  PubMed  Google Scholar 

  3. Acampora,D. et al. Forebrain and midbrain regions are deleted in Otx2-/- mutants due to a defective anterior neuroectoderm specification during gastrulation. Development 121, 3279–3290 (1995).

    CAS  PubMed  Google Scholar 

  4. Matsuo,I., Kuratani,S., Kimura,C., Takeda,N., Aizawa,S. Mouse Otx 2 functions in the formation and patterning of rostral head. Genes Dev. 9, 2646–2658 (1995).

    Article  CAS  PubMed  Google Scholar 

  5. Rhinn,M. et al. Sequential roles for Otx2 in visceral endoderm and neurectoderm for forebrain and midbrain induction and specification. Development 125, 845–856 (1998).

    CAS  PubMed  Google Scholar 

  6. Acampora,D. et al. Epilepsy and brain abnormalities in mice lacking Otx1 gene. Nature Genet. 14, 218–222 (1996).

    Article  CAS  PubMed  Google Scholar 

  7. Acampora,D., Avantaggiato,V., Tuorto,F. & Simeone,A. Genetic control of brain morphogenesis through Otx gene dosage requirement. Development 124, 3639–3650 (1997).

    CAS  PubMed  Google Scholar 

  8. Suda,Y., Matsuo,I. & Aizawa,S. Cooperation between Otx1 and Otx2 genes in developmental patterning of rostral brain. Mech. Dev. 69, 125–141 (1997).

    Article  CAS  PubMed  Google Scholar 

  9. Wassarmann,K. M. et al. Specification of the anterior hindbrain and establishment of a normal mid/hindbrain organizer is dependent on Gbx2 gene function. Development 124, 2923–2934 (1997).

    Google Scholar 

  10. Echelard,Y., Vassileva,G. & McMahon,A. P. Cis-acting regulatory sequences governing Wnt1 expression in the developing mouse CNS. Development 120, 2213–2224 (1994).

    CAS  PubMed  Google Scholar 

  11. Martinez,S., Marin,F., Nieto,M. A. & Puelles,L. Induction of ectopic engrailed expression and fat change in avian rhombomeres: intersegmental boundaries as barriers. Mech. Dev. 51, 289–303 (1995).

    Article  CAS  PubMed  Google Scholar 

  12. Hogan,B., Beddington,R., Constantini,F. Manipulating the Mouse Embryo. A Laboratory Manual (Cold Spring Harbor Laboratory, New York, 1994).

    Google Scholar 

  13. Wilkinson,D. In Situ Hybridization. A Practical Approach (IRL, Oxford, 1992).

    Google Scholar 

  14. Schaeren-Wiemers,N. & Gerfin-Moser,A. A single protocol to detect transcripts of various types and expression levels in neural tissue and cultured cells: in situ hybridization using digoxigenin-labelled cRNA probes. Histochemistry 100, 431–440 (1993).

    Article  CAS  PubMed  Google Scholar 

  15. McMahon,A. P., Joyner,A. L., Bradley,A. & McMahon,J. A. The midbrain-hindbrain phenotype of Wnt1-/Wnt1-mice results from stepwise deletion of engrailed-expressing cell by 9.5 days postcoitum. Cell 69, 581–595 (1992).

    Article  CAS  PubMed  Google Scholar 

  16. Millen,K. J., Hui,C. C. & Joyner,A. L. A role for En-2 and other murine homologues of Drosophila segment polarity genes in regulating positional information in the developing cerebellum. Development 121, 3935–3945 (1995).

    CAS  PubMed  Google Scholar 

  17. Frohman,M. A., Boyle,M. & Martin,G. Isolation of the mouse Hox-2.9 gene: analysis of embryonic expression suggests that positional information along the anterior posterior axis is specified by mesoderm. Development 110, 589–607 (1990).

    CAS  PubMed  Google Scholar 

  18. Toresson,H., Mata de Urquiza,A., Fragerstrom,C., Perlmann,T. & Campbell,K. Retinoids are produced by glia in the lateral ganglionic eminence and regulate striatal neuron differentiation. Development 126, 1317–1326 (1999).

    CAS  PubMed  Google Scholar 

  19. Puelles,L. & Rubenstein,J. L. R. Expression of homeobox and other putative regulatory genes in the mouse forebrain suggest a neuromeric organization. Trends Neurosci. 16, 472–479 (1993).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank A. Liu and Y. Li for helpful discussions and comments on the manuscript, and the Skirball transgenic facility for assistance with transgenic production. This work was supported by grants from the Human Frontiers Science Program (HFSP) and NINDS. S.M. was supported by an HFSP fellowship, and K.C. and D.J.E. by Canadian MRC fellowships. A.L.J. is an HHMI investigator.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Alexandra L. Joyner.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Millet, S., Campbell, K., Epstein, D. et al. A role for Gbx2 in repression of Otx2 and positioning the mid/hindbrain organizer. Nature 401, 161–164 (1999). https://doi.org/10.1038/43664

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/43664

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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