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Single-Cell Lineage Analysis of Oogenesis in Mice

  • Lei LeiEmail author
  • Allan C. SpradlingEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1463)

Abstract

Lineage analysis is widely used because it provides a very powerful tool for characterizing the developmental behavior of the cells in vivo. In this chapter, we describe a particularly informative variant of lineage analysis that we term “single-cell lineage analysis”. As in traditional lineage analysis, the method employs a Tamoxifen (Tmx)-inducible CAGCreER mouse line, which is crossed to an R26R reporter line that can be activated by Cre-mediated DNA recombination. However, instead of driving CreER at a high level within a subset of cells defined by a particular promoter, CreER is driven with a generic promoter that is active in essentially all cells throughout the lifespan of the mouse. Specificity comes from using only a very low dose of Tmx so that just a few random, widely separated individual cells undergo recombination and become labeled. The growth and behavior of most such initially marked cells can subsequently be followed over time because each one forms a growing clone of marked cells that does not overlap with other clones due to their rarity. Following individual cell growth patterns provides much more information than can be derived from traditional lineage analysis, which relies on promoter specificity and uses high doses of Tmx that cannot resolve the behavior of single cells. We illustrate the value of single-cell lineage analysis using a recent study of fetal germ cell development and a recent search for female germ-line stem cells in adult mouse ovaries.

Key words

Tamoxifen CreER Germ-line stem cells Ovary Oocyte Mouse 

Notes

Acknowledgement

This work was supported by Howard Hughes Medical Institute. We thank Dr. Matthew Sieber, Dr. Ethan Greenblatt, and Jui-Ko Chang for comments on the chapter.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Cell and Developmental BiologyUniversity of Michigan Medical SchoolAnn ArborUSA
  2. 2.Department of EmbryologyHoward Hughes Medical Institute, Carnegie Institution for ScienceBaltimoreUSA

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