Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Purification of GFRα1+ and GFRα1- spermatogonial stem cells reveals a niche-dependent mechanism for fate determination

ABSTRACT: Undifferentiated spermatogonia comprise a pool of stem cells and progenitor cells that show heterogeneous expression of markers, including the cell surface receptor GFRα1. Technical challenges in isolation of GFRα1+ versus GFRα1- undifferentiated spermatogonia have precluded the comparative molecular characterization of these subpopulations and their functional evaluation as stem cells. Here, we develop a method to purify these subpopulations by FACS and show that GFRα1+ and GFRα1- undifferentiated spermatogonia both demonstrate elevated transplantation activity while differing principally in receptor tyrosine kinase signaling and cell cycle. We identify the cell surface molecule MCAM as differentially expressed in these populations and show that antibodies to MCAM allow isolation of highly enriched populations of GFRα1+ and GFRα1- spermatogonia from adult, wild-type mice. In germ cell culture, GFRα1- cells upregulate MCAM expression in response to GDNF/FGF stimulation. In transplanted hosts, GFRα1- spermatogonia yield GFRα1+ spermatogonia and restore spermatogenesis, albeit at lower rates than their GFRα1+ counterparts. Together, these data provide support for a model of a stem cell pool in which the GFRα1+ and GFRα1- cells are closely related, but show key cell-intrinsic differences and can interconvert between the two states based, in part, on access to niche factors.

ORGANISM(S): Mus musculus

PROVIDER: GSE107694 | GEO | 2018/12/01

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Murine Spermatogonial Stem Cells: Rapamycin- vs Vehicle-exposed in vivo (2-wk treatment)

Project description:Male FVB strain mice aged 12-days-old through 26-days-old were administered daily intraperitoneal injections of rapamycin (4mg/kg body weight) or control vehicle (5% Tween-80, 5% PEG-400), beginning at postnatal day (P)12. Mice were euthanized at P26 and their testes were isolated for germ cell enrichment. Single cell suspensions of germ cells were prepared from isolated testes and subjected to magnetic-activated cell sorting (MACS). This procedure enriches the undifferentiated spermatogonia fraction, which represents the spermatogonial stem cell (SSC) population. Total RNA from cells double-positive for the SSC surface markers thymus cell antigen 1, theta (THY1) and glial cell line-derived neurotrophic factor family receptor alpha 1 (GFRA1) was isolated for gene expression microarray analysis. Magnetic-activated cell sorting (MACS) was used to enrich undifferentiated spermatogonia from a pool of primary testicular cells isolated from 5 littermate rapamycin (RM)-treated male mice and from 5 littermate control vehicle (VEH)-treated male mice. With MACS-enriched cells from pool generating RNA, no technical replicates performed.

2012-04-30 | E-GEOD-37062 | biostudies-arrayexpress

Cooperative polarization of MCAM/CD146 and ERM family proteins in melanoma

Project description:The WRAMP structure is a protein network associated with tail-end actomyosin contractility, membrane retraction, and directional persistence during cell migration. A marker of WRAMP structures is melanoma cell adhesion molecule (MCAM) which dynamically polarizes to the cell rear. However, factors that mediate MCAM polarization are still unknown. In this study, BioID using MCAM as bait identifies the ERM family proteins, moesin, ezrin, and radixin, as WRAMP structure components. We also present a novel image analysis pipeline, Protein Polarity by Percentile (“3P”), which classifies protein polarization using machine learning and facilitates quantitative analysis. Using 3P, we find that depletion of moesin, and to a lesser extent ezrin, decreases the proportion of cells with polarized MCAM. Furthermore, although co-polarized MCAM and ERM proteins show high spatial overlap, 3P identifies subpopulations with ERM proteins closer to the cell periphery. Live-cell imaging confirms that MCAM and ERM protein polarization is tightly coordinated, but ERM proteins enrich at the cell edge first. Finally, deletion of a juxtamembrane segment in MCAM previously shown to promote ERM protein interactions impedes MCAM polarization. Our findings highlight the requirement for ERM proteins in recruitment of MCAM to WRAMP structures and an advanced computational tool to characterize protein polarization.

2024-05-22 | PXD047722 | Pride

Progression of spermatogonial differentiation in the mouse: the pathway to meiosis

Project description:We report bulk RNA-sequencing data for undifferentiated and defined subpopulations of differentiating murine spermatogonia. We have sequenced two replicates each of wild-type and STRA8 knock-out sets of spermatogonia to assess transcriptional changes that occur throughout spermatogonial development prior to meiotic entry.

2020-07-29 | GSE155307 | GEO

Mus musculus

Project description:RNA-seq analyses of ID4-EGFP+ undifferentiated spermatogonia and sorted subpopulations.

| PRJNA325534 | ENA

Analysis of gene expression heteroegeneity in adult undifferentiated spermatogonia [single-cell RNA-Seq]

Project description:The undifferentiated spermatogonial population of mouse testis is functionally heterogeneous and contains stem cells and committed progenitor cells. However, gene expression patterns marking these distinct cell fractions are poorly defined. Therefore, our aim was to profile gene expression of undifferentiated cells by RNA-Seq at the single cell level. Undifferentiated cells were therefore isolated from adult mouse testes based on expression of a Plzf/Zbtb16 gene reporter, which marks both stem and progenitor spermatogonia. Isolated cells were then processed using the 10X Chromium system and analysed by RNA-Seq. Our goal was to characterize distinct subsets of undifferentiated spermatogonia based on gene expression patterns.

2018-07-26 | GSE112880 | GEO

Identification of gene targets of the transcription factor SALL4 in undifferentiated spermatogonia

Project description:Sustained spermatogenesis in adult males and recovery of fertility following germ cell depletion are dependent on undifferentiated spermatogonia with self-renewal potential. We have previously demonstrated a critical role for the transcription factor Spalt-like 4 (SALL4) in spermatogonial differentiation. However, it remains unclear whether SALL4 has broader roles within the spermatogonial pool despite its ability to co-regulate genes with PLZF, a transcription factor required for undifferentiated cell maintenance. To identify genes regulated by SALL4 in the male germline, we established cultures of undifferentiated spermatogonia from a Sall4 inducible knockout mouse model. Cells were treated with vehicle (as control) or tamoxifen to induce gene deletion, then cells harvested and analysed by microarray to identify genes mis-expressed upon loss of SALL4.

2017-08-31 | GSE98991 | GEO

A bioenergetic shift is required for spermatogonial differentiation

Project description:A bioenergetic balance between glycolysis and mitochondrial respiration is particularly important for stem cell fate specification. It however remains to be determined whether undifferentiated spermatogonia switch their preference of bioenergy production during differentiation. In this study, we found that ATP generation in spermatogonia was gradually increased upon retinoic acid-induced differentiation. To accommodate this elevated energy demand, retinoic acid signaling concomitantly switched ATP production in spermatogonia from glycolysis to mitochondrial respiration, accompanied by increased levels of reactive oxygen species. In addition, inhibition of glucose conversion to glucose-6-phosphate or pentose phosphate pathway blocked the formation of c-Kit+ differentiating germ cells, suggesting that metabolites produced from glycolysis are required for spermatogonial differentiation. We further demonstrated that the expression levels of several metabolic regulators and enzymes were significantly altered upon retinoic acid-induced differentiation by both RNA-seq analyses and quantitative proteomics. Taken together, our data unveil a critically regulated bioenergetic balance between glycolysis and mitochondrial respiration which is required for spermatogonial proliferation and differentiation.

2021-09-09 | PXD019136 | Pride

LNGFR- and Thy-1-based prospective isolation of human mesenchymal stem cells reveals functionally distinct subpopulations.

Project description:Human multipotent mesenchymal stem cells (MSCs), isolated based on their adherence to plastic, show poor growth and differentiation and frequently contain contaminating cells, which limits to clarify their own characteristics. In this study, the identification of two cell surface markers, LNGFR and Thy-1, allowed the prospective isolation of highly purified clonogenic human MSCs. Furthermore, single cell sorting assays followed by in vitro expansion revealed three distinct MSC subpopulations: rapidly-, moderately- and slowly proliferating MSC clones (RPCs, MPCs and SPCs, respectively). While RPCs exhibited robust multilineage differentiation and self-renewal potency, MPCs and SPCs contained a majority of senescent cells and exhibited frequent genome errors. Single cell sorting assays followed by in vitro expansion revealed three distinct MSC subpopulations: rapidly-, moderately- and slowly proliferating MSC clones (RPCs, MPCs and SPCs, respectively). Sample: RPC, MPC and SPC (n=3).

2011-12-22 | E-GEOD-34484 | biostudies-arrayexpress

Differentially expressed genes in undifferentiated spermatogonia at single cell level following a low dose of busulfan

Project description:Spermatogonial stem cells are responsible for sustaining gametes production and male fertiliy in men. However, germ cells including SSCs are highly sensitive to chemotherapies and radiations, placing male cancer patients at high risk of treatment-induce infertility. We have previously shown that the undifferentiated spermatogonial population of mouse testis is functionally heterogeneous and contain both stem cells and committed progenitor cells. Using mouse model of chemotherapy-induced germline damage and recovery, we aim to define molecular charactersitics, dynamics and heterogeneity of undifferentiated spermatogonia during germline regeneration compared to homeostatic undifferentiated spermatogonia.

2022-03-04 | GSE182924 | GEO

PLZF and DDX5 interacting proteins in spermatogonia

Project description:DDX5, or PLZF co-immunoprecipitation in lysates from cultured undifferentiated spermatogonia followed by identification of eluted proteins using mass spectrometry. IgG control IPs included.

2019-02-24 | MSV000083476 | MassIVE

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data