Summary
Although a common approach in large vertebrate embryos such as chick or frog, manipulation at the tissue level is only rarely applied to zebrafish embryos. Despite its relatively small size, the zebrafish embryo can be readily used for micromanipulations such as tissue and organ primordium transplantation, explantation, and microbead implantation, to study inductive tissue interactions and tissue autonomy of pleiotropic, mutant phenotypes or to isolate tissue for organotypic and primary cell culture or RNA isolation. Since this requires special handling techniques, tools, and tricks, which are rarely published and thus difficult to apply without hands-on demonstration, this article provides detailed instructions and protocols on tissue micromanipulation. The goal is to introduce a broader scientific audience to these surgical techniques, which can be applied to a wide range of questions and used as a starting point for many downstream applications in the genetically tractable zebrafish embryo.
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Spemann, H. and Mangold, H. (1924). Über Induktion von Embryonanlagen durch Implantation artfremder Organisatoren. Arch. Mikr. Anat. Entw. Mech. 100, 599–638.
Reifers, F., Walsh, E.C., Lger, S., Stainier, D.Y. and Brand, M. (2000). Induction and differentiation of the zebrafish heart requires fibroblast growth factor 8 (fgf8/acerebellar). Development 127, 225–235.
Picker, A. and Brand, M. (2005). Fgf signals from a novel signaling center determine axial patterning of the prospective neural retina. Development 132, 4951–4962.
Jászai, J., Reifers, F., Picker, A., Langenberg, T. and Brand, M. (2003). Isthmus-to-midbrain transformation in the absence of midbrainhindbrain organizer activity. Development 130, 6611–6623.
Scholpp, S. and Brand, M. (2004). Endocytosis controls spreading and effective signaling range of Fgf8 protein. Curr Biol. 14, 1834–1841.
von der Hardt, S., Bakkers, J., Inbal, A., Carvalho, L., Solnica-Krezel, L., Heisenberg, C.P., et al (2007). The Bmp gradient of the zebrafish gastrula guides migrating lateral cells by regulating cell-cell adhesion. Curr Biol. 17, 475–487.
Picker, A., Brennan, C., Reifers, F., Clarke, J.D., Holder, N. and Brand, M. (1999). Requirement for the zebrafish mid-hindbrain boundary in midbrain polarisation,mapping and confinement of the retinotectal projection. Development 126, 2967–2978.
Wagle, M., Grunewald, B., Subburaju, S., Barzaghi, C., le Guyader, S., Chan, J., et al (2004). EphrinB2a in the zebrafish retinotectal system. J. Neurobiol. 59, 57–65.
Fricke, C., Lee, J.S., Geiger-Rudolph, S., Bonhoeffer, F. and Chien, C.B. (2001). Astray, a zebrafish roundabout homolog required for retinal axon guidance. Science 292, 507–510.
Hutson, L.D., Campbell, D.S. and Chien, C.B. (2004). Analyzing axon guidance in the zebrafish retinotectal system. Methods Cell Biol 76, 13–35.
Hutson, L.D., Campbell, D.S. and Chien, C.B. (2004). Analyzing axon guidance in the zebrafish retinotectal system, in Methods in Cell Biology, Volume 76, The Zebrafish: cellular and Developmental Biology (Detrich, H.W., III., Westerfield, M., Zon, L.I., eds.), Elsevier Academic Press, London, pp. 13–32.
Kane, D.A. and Kishimoto, Y. (2002). Cell labelling and transplantation techniques, in Zebrafish: A Practical Approach (Nuesslein-Volhard, C., Dahm, R., eds.), Oxford University Press, Oxford, pp. 95–119).
Detrich, H.W., III., Westerfield, M. and Zon, L.I. (eds.) (2004). Methods in Cell Biology, Volume 76, The Zebrafish: Cellular and Developmental Biology, Elsevier Academic Press, London.
Westerfield, M. (2000). The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio). 4th ed., University of Oregon Press, Eugene.
Langenberg, T., Brand, M. and Cooper, M.S. (2003). Imaging brain development and organogenesis in zebrafish using immobilized embryonic explants. Dev Dyn. 228, 464–474.
Dquant, M.L., Glynn, E., Gaudenz, K., Wahl, M., Chen, J., Mushegian, A., et al (2006). A complex oscillating network of signaling genes underlies the mouse segmentation clock. Science 314, 1595–1598.
Acknowledgments
This work was supported by grants to MB from the Deutsche Forschungsgemeinschaft (SFB 655), and the EU (ZF-Models and Endotrack) and by the Max Planck Society. The authors would like to thank Mary-Lee Dequeant, Aurelie Krol, Alexander Aulehla, and Karen Echeverri for advice on PSM dissection and technical support, the fish facilities of the MPI-CBG and Biotechnology Center, Dresden and Stowers Institute, Kansas City, MO, USA, and Katrin Bergman (CRTD) for photography.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Picker, A., Roellig, D., Pourquié, O., Oates, A.C., Brand, M. (2009). Tissue Micromanipulation in Zebrafish Embryos. In: Lieschke, G., Oates, A., Kawakami, K. (eds) Zebrafish. Methods in Molecular Biology, vol 546. Humana Press. https://doi.org/10.1007/978-1-60327-977-2_11
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DOI: https://doi.org/10.1007/978-1-60327-977-2_11
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