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17 - Retinoic acid signalling controls anteroposterior patterning of the zebrafish hindbrain

Published online by Cambridge University Press:  11 August 2009

G. Begemann
Affiliation:
Department of Biology, University of Konstanz, Universitaetsstr. 10, D-78464 Konstanz, Germany
Manuel Marí-Beffa
Affiliation:
Universidad de Málaga, Spain
Jennifer Knight
Affiliation:
University of Colorado, Boulder
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Summary

OBJECTIVE OF THE EXPERIMENT The zebrafish, Danio rerio, is a successful addition to the collection of vertebrate model systems that offers several attractive features: mating pairs are readily available and spawn large numbers of eggs on a regular basis. Development is rapid and occurs entirely outside the mother, and embryos and larvae are completely transparent, allowing the observation of the circulatory system and internal organs in the living organism. Up to a few thousand mutants have now been isolated through mutagenesis screens that allow a systematic dissection of vertebrate developmental genetics, mirroring the success of screens in Drosophila and C. elegans.

Segmentation of the body along the anteroposterior axis is a feature found in arthropods, annelids and chordates. In the zebrafish, as in other vertebrates, segmentation is apparent at embryonic stages in the subdivision of the trunk paraxial mesoderm into somites. Although less obvious, the anterior neural tube is also segmentally arranged, as can be seen in the partition of the prospective hindbrain, the rhombencephalon, into seven rhombomeres.

Here the mechanisms that establish anteroposterior identity among rhombomeres will be analysed at the gene expression level. In particular, the effects of exogenous manipulation of retinoic acid (RA) signalling on the development of the central nervous system will be studied in the zebrafish embryo.

DEGREE OF DIFFICULTY Experiments require breeding pairs of zebrafish and some competence in handling fish and setting up pair matings. Both experiments require basic molecular techniques and care in minimising RNAse contamination.

Type
Chapter
Information
Publisher: Cambridge University Press
Print publication year: 2005

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References

Begemann, G., Schilling, T. F., Rauch, G. J., Geisler, R., and Ingham, P. W. (2001). The zebrafish neckless mutation reveals a requirement for raldh2 in mesodermal signals that pattern the hindbrain. Development, 128, 3081–94Google ScholarPubMed
Begemann, G., Marx, M., Mebus, K., Meyer, A., and Bastmeyer, M. (2004). Beyond the neckless phenotype: Influence of reduced retinoic acid signaling on motorneuron development in the zebrafish hindbrain. Dev. Biol., 271, in pressCrossRef
Grandel, H., Lun, K., Rauch, G. J., Rhinn, M., Piotrowski, T., Houart, C., Sordino, P., Kuchler, A. M., Schulte-Merker, S., Geisler, R., Holder, N., Wilson, S. W., and Brand, M. (2002). Retinoic acid signalling in the zebrafish embryo is necessary during pre-segmentation stages to pattern the anterior–posterior axis of the CNS and to induce a pectoral fin bud. Development, 129, 2851–65Google Scholar
Jowett, T. (1997). Tissue “in Situ” Hybridization: Methods in Animal Development. New York: John Wiley and Sons
Lumsden, A., and Krumlauf, R. (1996). Patterning the vertebrate neuraxis. Science, 274, 1109–15CrossRefGoogle ScholarPubMed
Maden, M. (2002). Retinoid signalling in the development of the central nervous system. Nature Reviews Neuroscience, 3, 843–53CrossRefGoogle ScholarPubMed
Nüsslein-Volhard, C., and Dahm, R. (2002). Zebrafish, A Practical Approach. Oxford: Oxford University Press
Westerfield, M. (1995). The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Danio rerio). Oregon: University of Oregon Press

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