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Biol Reprod. 2016 Dec;95(6):117. Epub 2016 Oct 12.

TSPAN8 Expression Distinguishes Spermatogonial Stem Cells in the Prepubertal Mouse Testis.

Author information

  • 1Department of Biology, University of Texas at San Antonio, San Antonio, Texas.
  • 2Genomics Core Facility, University of Texas at San Antonio, San Antonio, Texas.
  • 3Center for Reproductive Biology, School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, Washington.
  • 4Department of Anatomy and Cell Biology and East Carolina Diabetes and Obesity Institute, Brody School of Medicine, East Carolina University, Greenville, North Carolina.
  • 5Department of Biology, University of Texas at San Antonio, San Antonio, Texas brian.hermann@utsa.edu.

Abstract

Precise separation of spermatogonial stem cells (SSCs) from progenitor spermatogonia that lack stem cell activity and are committed to differentiation remains a challenge. To distinguish between these spermatogonial subtypes, we identified genes that exhibited bimodal mRNA levels at the single-cell level among undifferentiated spermatogonia from Postnatal Day 6 mouse testes, including Tspan8, Epha2, and Pvr, each of which encode cell surface proteins useful for cell selection. Transplantation studies provided definitive evidence that a TSPAN8-high subpopulation is enriched for SSCs. RNA-seq analyses identified genes differentially expressed between TSPAN8-high and -low subpopulations that clustered into multiple biological pathways potentially involved in SSC renewal or differentiation, respectively. Methyl-seq analysis identified hypomethylated domains in the promoters of these genes in both subpopulations that colocalized with peaks of histone modifications defined by ChIP-seq analysis. Taken together, these results demonstrate functional heterogeneity among mouse undifferentiated spermatogonia and point to key biological characteristics that distinguish SSCs from progenitor spermatogonia.

KEYWORDS:

epigenetics; single-cell; spermatogonial stem cells; surface marker; transcriptome; transplantation

PMID:
27733379
DOI:
10.1095/biolreprod.116.144220
[PubMed - in process]
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