Project description:YTHDF2 displays extensive-expression patterns during oocyte maturation and its deficiency causes female infertility in mice. However, its specific mechanism of regulation remains elusive due to the absence of suitable in vitro models. FGSCs possess the capacity for self-renewal and differentiation into oocytes to support reproduction. The successful establishment of a line of FGSCs provides a platform for scientific research on female fertility and oogenesis. To understand how YTHDF2 exerts its regulatory effects on FGSCs, we conducted MeRIP-seq assay in FGSCs and aimed to identified YTHDF2 target transcripts.
Project description:Metformin increase the number of FGSCs in mouse ovary, and the increased pan-β-hydroxylbutylated protein modification level in vitro FGSCs was detected by WB. There are the Mass spectrum original data of proteomic examination about pan-β-hydroxybutyrylation sites in female germline stem cells after merformin treatment
Project description:Alternative polyadenylation (APA) is an important post-transcriptional modification implicated in development. Female germline stem cell (FGSC) is unipotent and capable of giving rise to oocyte. However, whether alternative polyadenylation plays a role in self-renew and cell fate determination of FGSCs remain elusive. Here, we used 3T-Seq developed in our lab to profile genome-wide 3a termini of transcripts and delineate APA sites in mouse FGSCs and explored the biological significance of APA modulation in FGSC identity.
Project description:YTHDF2 displays extensive-expression patterns during oocyte maturation and its deficiency causes female infertility in mice. However, its specific mechanism of regulation remains elusive due to the absence of suitable in vitro models. Female germline stem cells (FGSCs) possess the capacity for self-renewal and differentiation into oocytes to support reproduction. The successful establishment of a line of FGSCs provides a platform for scientific research on female fertility and oogenesis. To understand how YTHDF2 exerts its regulatory effects on FGSCs, we conducted RNA-seq assay in WT and Ythdf2-KO FGSCs and aimed to identified YTHDF2-responsive genes.
Project description:Female germline stem cells (FGSCs) are adult stem cells capable of self-renewal and differentiation into mature oocytes. AKT3, a member of the AKT kinase family, plays crucial roles in multiple cellular processes, such as proliferation, migration, and apoptosis. However, the mechanism by which AKT3 affects the development of FGSCs is poorly understood. We performed liquid chromatography (LC)-mass spectrometry (MS) on mouse FGSCs in which AKT3 was knocked down using a lentivirus and on control FGSCs to investigate how AKT affects the development of FGSCs. Based on the raw LC-MS data and database searches and data filtering, we identified 46,260 peptides, including 45,821 unique peptides, corresponding to 6849 identified proteins and 6697 comparable proteins. These identified proteins were functionally annotated using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Protein Domain, Clusters of Orthologous Genes (COG)/Eukaryotic Orthologous Groups (KOG), STRING database, Reactome, WikiPathways, HallMark, and transcription factor (TF) analyses. Fisher’s exact test was used to assess the significance of functional enrichment of the differentially abundant proteins. We identified 281 differentially abundant proteins between AKT3 knockdown and control FGSCs, comprising 229 upregulated and 52 downregulated proteins. We performed clustering analysis on these differentially abundant proteins based on functional enrichment using GO, Domain, KEGG, Reactome and WikiPathways platforms. A protein–protein interaction network was constructed to demonstrate interactions between proteins. These datasets will facilitate future investigations into the mechanisms governing FGSC self-renewal and differentiation and will provide a foundation for understanding diseases related to abnormal germ cell development.
Project description:Germ stem cells are the only stem cells which can produce gametes in vivo. Recently, accumulating evidence demonstrates the isolation and culture of female germline stem cells (FGSCs) from adult mice and humans. Here, we established FGSCs line from single EGFP transgenetic mouse, and traced their differentiation in vivo. Then we compared the follile development from transplanted FGSCs mice (F-TF) with that from wide type mice (F-WT) by RNA-Seq. qRT–PCR validation was performed using SYBR Green assays. Using Illumina HiSeq2500 sequencer, we generated 51.5 Gb of sequencing data from the 8 samples. On average, we detected expression of 20 966 transcripts. Under the criteria fold change > 2 or < 0.5, we obtained 7005 and 5764 differential expressed genes (DEGs) from F-TF or F-WT,respectively. Weighted correlation network analysis (WGCNA) analysis identified core genes and two potential networks during folliculogenesis which were mainly down-regulated after preantral follicle. This study was the first to analyze the development of transplanted FGSCs progressively and would provide the theoretical basis for clinical translational research of FGSCs.
Project description:We used the microarray analysis to compare the globe gene expression profiles of female germ line stem cells (FGSCs) and spermatogonial stem cells (SSCs).