Single-cell lineage analysis of oogenesis in mice

Lei Lei, Allan C. Spradling

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages125-138
Number of pages14
Volume1463
DOIs
StatePublished - 2017
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1463
ISSN (Print)10643745

Keywords

  • CreER
  • Germ-line stem cells
  • Mouse
  • Oocyte
  • Ovary
  • Tamoxifen

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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