Abstract
Microinjection remains the most popular and effective of the methods to introduce DNA, RNA, and proteins into fertilized zebrafish eggs. The method is simple and reliable. A microinjection pipet is filled with the DNA or RNA solution and attached to an apparatus that forces the solution out of the pipet with air pressure. A small amount of solution is then expelled into the cytoplasm of the embryo before withdrawing the pipet, and the injected embryos are incubated to develop further. Once inside the cells, the foreign DNA or RNA is transcribed and/or translated within the developing embryos and the functional roles of their protein products can be evaluated by morphological, physiological or molecular changes. Thus, microinjection has been widely used for generating transgenic fish (1–3), analyzing gene function by overexpression of DNA or RNA (4–6) and mapping cell fate in early blastula embryos (7,8).
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Stuart, G. W., McMurray, J. V., and Westerfield, M. (1988) Replication, integration and stable germ-line transmission of foreign sequences injected into early zebrafish embryos. Development 103, 403–412.
Stuart, G. W., Vielkind, J. R., McMurray, J. V., and Westerfield, M. (1990) Stable lines of transgenic zebrafish exhibit reproducible patterns of transgene expression. Development 109, 557–584.
Culp, P., Nusslein-Volhard, C., and Hopkins, N. (1991) High frequency germ-line transmission of plasmid DNA sequences injected into fertilised zebrafish eggs. Proc. Natl. Acad. Sci. USA 88, 7953–7957.
Kelly, G. M., Greenstein, P., Erezyilmaz, D. F., and Moon, R. T. (1995) Zebrafish Wnt8 and Wnt 8b share a common activity but are involved in distinct developmental pathways. Development 121, 1787–1799.
Xu, Q. L., Alldus, G., Holder, N., and Wilkinson, D. G. (1995) Expression of truncated Sek-1 receptor tyrosine kinase disrupts the segmental restriction of gene-expression in the Xenopus and zebrafish hindbrain. Development 121, 4005–4016.
Long, Q. M., Meng, A. M., Wang, H., Jessen, J. R., Farrell, M. J., and Lin, S. (1997) GATA-1 expression pattern can be recapitulated in living transgenic zebrafish using GFP reporter gene. Development 124, 4105–4111.
Strehlow, D., Heinrich, G., and Gilbert, W. (1994) The fates of the blastomeres of the 16-cell zebrafish embryo. Development 120, 1791–1798.
Helde, K. A., Wilson, E. T., Cretekos, C. J., and Grunwald, D. J. (1994) Contribution of early cells to the fate map of the zebrafish gastrula. Science 265, 517–520.
Lin, S., Gaiano, N., Culp, P., Burns, J. C., Friedmann, T., Yee, J.-K., et al. (1994) Integration and germ-line transmission of a pseudotyped retroviral vector in zebrafish. Science 265, 666–669.
Gaiano, N., Allende, M., Amsterdam, A., Kawakami, K., and Hopkins, N. (1996) Highly efficient germ-line transmission of proviral insertions in zebrafish. Proc. Natl. Acad. Sci. USA 93, 7777–7782.
Westerfield, M. (1995) The Zebrafish Book, A Guide for the Laboratory Use of Zebrafish, 3rd ed., University of Oregon Press, Eugene, OR.
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© 1999 Humana Press Inc.
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Xu, Q. (1999). Microinjection into Zebrafish Embryos. In: Guille, M. (eds) Molecular Methods in Developmental Biology. Methods in Molecular Biology™, vol 127. Humana Press. https://doi.org/10.1385/1-59259-678-9:125
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DOI: https://doi.org/10.1385/1-59259-678-9:125
Publisher Name: Humana Press
Print ISBN: 978-0-89603-790-8
Online ISBN: 978-1-59259-678-2
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