Cleavage Blastomere Deletion and Transplantation to Test Cell Fate Commitment in Xenopus

Cold Spring Harbor Protocols
S A Moody

Abstract

Fate maps identify the precursors of an organ, and tracing the members of a blastomere lineage over time shows how its descendants come to populate that organ. The fates of the individual blastomeres of the two- to 32-cell Xenopus embryo have been fully mapped to reveal which cells are the major contributors to various cell types, tissues, and organs. However, because these fate maps were produced in the normal embryo, they do not reveal whether a precursor blastomere is competent to give rise to additional tissues or is already committed to its fate-mapped repertoire of descendants. To identify the mechanisms by which a cell's fate is committed, one needs to expose the cell to different experimental environments. If the cell's fate is determined, it will express its normal fate or gene expression profile in novel environments, whereas if it is not yet determined it will express different fates or gene expression profiles when exposed to novel external factors or neighboring cells. This protocol describes two techniques for testing cell fate commitment: single cell deletion and single cell transplantation. Deleting a blastomere allows one to test whether the deleted cell is required for the remaining cells to produce their norm...Continue Reading

References

Jan 1, 1990·Anatomy and Embryology·S A Moody, M J Kline
Aug 1, 1987·Developmental Biology·S A Moody
Feb 1, 1987·Developmental Biology·S A Moody
Jan 1, 2007·CSH Protocols·Hazel L SiveRichard M Harland
Jan 1, 2007·CSH Protocols·Hazel L SiveRichard M Harland
Nov 1, 2017·Cold Spring Harbor Protocols·Jean-Pierre Saint-Jeannet
Dec 2, 2017·Cold Spring Harbor Protocols·Michael W Klymkowsky
May 18, 2018·Cold Spring Harbor Protocols·Sally A Moody
May 18, 2018·Cold Spring Harbor Protocols·Sally A Moody
May 18, 2018·Cold Spring Harbor Protocols·Sally A Moody

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Citations

May 18, 2018·Cold Spring Harbor Protocols·Sally A Moody
May 18, 2018·Cold Spring Harbor Protocols·Sally A Moody
Jun 13, 2020·Frontiers in Cellular Neuroscience·Vaibhav P PaiMichael Levin

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