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24 - The conservation of the genome and nuclear reprogramming in Xenopus

Published online by Cambridge University Press:  11 August 2009

J. B. Gurdon
Affiliation:
Wellcome Trust/CRC Cancer UK Institute, Institute of Cancer and Developmental Biology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK
Manuel Marí-Beffa
Affiliation:
Universidad de Málaga, Spain
Jennifer Knight
Affiliation:
University of Colorado, Boulder
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Summary

OBJECTIVE OF THE EXPERIMENT A fundamental question in developmental biology is whether the processes of development and cell differentiation involve a stable change in the genome of cells, as was thought to be the case many years ago. An alternative idea is that the genome remains constant in cells of all different types and that the readout of the genome, namely transcription and translation, is modified according to cell type.

The first experiments to address this question and to provide most of an answer to it were carried out with amphibian eggs and embryos, using the technique of nuclear transplantation. Amphibian eggs have provided the most favoured material for embryological research since the late 1800s until the last two or three decades. The classical embryological experiments from Spemann onwards were conducted with this material. Nuclear transplantation exemplifies the value of amphibian eggs for embryological experiments involving manipulation of embryos and cells.

The aim of the experiment described below is to test the proposition that cells can embark on a particular pathway of differentiation while retaining a complete, totipotent, or at least multipotent, genome in their cells.

Further technical details required for this experiment may be found in Gurdon, 1991.

DEGREE OF DIFFICULTY From an intellectual point of view the experiment is extremely simple. It aims to replace the zygote nucleus of the fertilized egg, i.e. the egg and sperm pronuclei, with the nucleus of a cell that has clearly embarked on one direction of differentiation.

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Publisher: Cambridge University Press
Print publication year: 2005

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References

Briggs, R., and King, T. J. (1952). Transplantation of living nuclei from blastula cells into enucleated frogs' eggs. Proc. Natl Acad. Sci. U.S.A. 38, 455–63CrossRefGoogle ScholarPubMed
Gurdon, J. B. (1960). The developmental capacity of nuclei taken from differentiating endoderm cells of Xenopus laevis. J. Embryol. Exp. Morphol. 8, 505–26Google ScholarPubMed
Gurdon, J. B. (1962). The developmental capacity of nuclei taken from intestinal epithelium cells of feeding tadpoles. J. Embryol. Exp. Morphol. 10, 622–40Google ScholarPubMed
Gurdon, J. B. (1977). Methods for nuclear transplantation in Amphibia. In Methods in Cell Biology 16, eds. G. Stein, J. Stein and K. J. Kleinsmith, pp. 125–39. New York: Academic Press
Gurdon, J. B. (1991). Nuclear transplantation in Xenopus. In Methods in Cell Biology 36, eds. B. K. Kay and H. B. Peng, pp. 299–309. San Diego: Academic Press
Gurdon, J. B., and Uehlinger, V. (1966). “Fertile” intestine nuclei. Nature 210, 1240–1CrossRefGoogle ScholarPubMed
Slack, J. M. W., and Forman, D. (1980). An interaction between dorsal and ventral regions of the marginal zone in early amphibian embryos. J. Embryol. Exp. Morphol. 56, 283–99Google ScholarPubMed

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