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Analysis of the C. elegans Germline Stem Cell Pool

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Germline Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2677))

Abstract

The Caenorhabditis elegans germline is an excellent model for studying the genetic and molecular regulation of stem cell self-renewal and progression of cells from a stem cell state to a differentiated state. The germline tissue is organized in an assembly line with the germline stem cell (GSC) pool at one end and differentiated gametes at the other. A simple mesenchymal niche caps the GSC pool and maintains GSCs in an undifferentiated state by signaling through the conserved Notch pathway. Notch signaling activates transcription of the key GSC regulators lst-1 and sygl-1 proteins in a gradient through the GSC pool. LST-1 and SYGL-1 proteins work with PUF RNA regulators in a self-renewal hub to maintain the GSC pool. In this chapter, we present methods for characterizing the C. elegans GSC pool and early stages of germ cell differentiation. The methods include examination of germlines in living and fixed worms, cell cycle analysis, and analysis of markers. We also discuss assays to separate mutant phenotypes that affect the stem cell vs. differentiation decision from those that affect germ cell processes more generally.

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Authors and Affiliations

  1. Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA

    Sarah L. Crittenden & Judith Kimble

  2. Department of Biology, Eastern Michigan University, Ypsilanti, MI, USA

    Hannah S. Seidel

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  1. Sarah L. Crittenden
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  2. Hannah S. Seidel
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  3. Judith Kimble
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Correspondence to Sarah L. Crittenden .

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  1. Department of Molecular Biology, The University of Texas Southwestern Medical Center, Dallas, TX, USA

    Michael Buszczak

1 Electronic Supplementary Material

Z-stack image of an adult hermaphrodite germline stained with DAPI to visualize DNA (magenta) and labeled with a 15-minute pulse of EdU to mark S-phase cells (green). Examples of cells in each stage of the cell cycle are marked. Note that G1 cells occur in pairs (MP4 684 kb)

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Crittenden, S.L., Seidel, H.S., Kimble, J. (2023). Analysis of the C. elegans Germline Stem Cell Pool. In: Buszczak, M. (eds) Germline Stem Cells. Methods in Molecular Biology, vol 2677. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3259-8_1

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