title: TSPAN8 Expression Distinguishes Spermatogonial Stem Cells in the Prepubertal Mouse Testis.
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.
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: Kazadi
authors.initials: K
authors.lastname: Mutoji
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: Anukriti
authors.initials: A
authors.lastname: Singh
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: Thu
authors.initials: T
authors.lastname: Nguyen
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas.|Genomics Core Facility, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: Heidi
authors.initials: H
authors.lastname: Gildersleeve
authors.affiliation: Center for Reproductive Biology, School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, Washington.
authors.forename: Amy V
authors.initials: AV
authors.lastname: Kaucher
authors.affiliation: Center for Reproductive Biology, School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, Washington.
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Oatley
authors.affiliation: Center for Reproductive Biology, School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, Washington.
authors.forename: Jon M
authors.initials: JM
authors.lastname: Oatley
authors.affiliation: Department of Anatomy and Cell Biology and East Carolina Diabetes and Obesity Institute, Brody School of Medicine, East Carolina University, Greenville, North Carolina.
authors.forename: Ellen K
authors.initials: EK
authors.lastname: Velte
authors.affiliation: Department of Anatomy and Cell Biology and East Carolina Diabetes and Obesity Institute, Brody School of Medicine, East Carolina University, Greenville, North Carolina.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Geyer
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: Keren
authors.initials: K
authors.lastname: Cheng
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: John R
authors.initials: JR
authors.lastname: McCarrey
authors.affiliation: Department of Biology, University of Texas at San Antonio, San Antonio, Texas brian.hermann@utsa.edu.|Genomics Core Facility, University of Texas at San Antonio, San Antonio, Texas.
authors.forename: Brian P
authors.initials: BP
authors.lastname: Hermann
mesh_terms: D000072956:Adult Germline Stem Cells / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D020869:Gene Expression Profiling; D008297:Male; D051379:Mice; D036104:Receptor, EphA2 / Q000235:genetics / Q000378:metabolism; D013091:Spermatogenesis; D013737:Testis / Q000166:cytology* / Q000378:metabolism; D059470:Tetraspanins / Q000235:genetics / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D059470:Tetraspanins; D036104:Receptor, EphA2
keywords: epigenetics; single-cell; spermatogonial stem cells; surface marker; transcriptome; transplantation