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MRNA-Seq profiling of Stra8-deficient and wild-type testes

ABSTRACT: We sequenced mRNA in grossly enlarged testes from 1-year-old Stra8-deficient mice, and in testes from adult male wild-type controls, to verify that Stra8-deficient testes are enriched for genes normally expressed in type A spermatogonia. Examination of mRNA levels in 6 whole-testis samples (3 replicates of each genotype). Specifically, we sequenced mRNA in grossly enlarged testes from three 1-year-old Stra8-deficient male mice, and in normal testes from three adult male wild-type controls

ORGANISM(S): Mus musculus

SUBMITTER: David Page

PROVIDER: E-GEOD-67169 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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mRNA-Seq profiling of Stra8-deficient and wild-type testes

Project description:We sequenced mRNA in grossly enlarged testes from 1-year-old Stra8-deficient mice, and in testes from adult male wild-type controls, to verify that Stra8-deficient testes are enriched for genes normally expressed in type A spermatogonia.

2015-04-03 | GSE67169 | GEO

The deubiquitinase cofactor UAF1 interacts with USP1 and plays an essential role in spermiogenesis

Project description:Spermiogenesis defines the final phase of male germ cell differentiation. Multiple deubiquitinating enzymes have been linked to spermiogenesis, yet the impacts of deubiquitination on spermiogenesis remain poorly characterized. Here, we investigated the function of UAF1 in mouse spermiogenesis and male fertility. We selectively deleted Uaf1 in premeiotic germ cells using Stra8-Cre knock-in mouse strain (Uaf1 sKO), and found that Uaf1 is essential for spermiogenesis and male fertility. We found that UAF1 interacts and colocalizes with USP1 in testes. Conditional knockout of Uaf1 in testes results in disturbed expression and localization of USP1, suggesting that UAF1 regulates spermiogenesis through the function of the deubiquitinating enzyme USP1. We used tandem mass tag-based proteomics to identify differentially expressed proteins and potential underlying mechanisms, and found that conditional knockout Uaf1 in testes results in reduced levels of proteins essential for spermiogenesis. Thus, the UAF1/USP1 deubiquitinase complex is essential for normal spermiogenesis by regulating the levels of spermiogenesis-related proteins.

2024-02-07 | PXD046120 | Pride

MIWI catalysis is required for piRNA amplification-independent LINE1 transposon silencing [microarray]

Project description:Here we show that MIWI is a small RNA-guided ribonuclease (Slicer) that requires extensive complementarity for target cleavage in vitro. Disruption of its catalytic activity in mice by a single point mutation results in male infertility and displays increased accumulation of LINE1 transposon transcripts. Round spermatids were purified from adult mouse testes of indicated Miwi genotypes. Total RNA was extracted and subjected to microarray analysis for gene expression profiling.

2011-11-30 | E-GEOD-32180 | biostudies-arrayexpress

Mus musculus

Project description:mRNA-Seq profiling of Stra8-deficient and wild-type testes

| PRJNA279154 | ENA

Global gene expression comparation among testes, ovaries, Smad4flox/floxMerCreMer Stra8+/− ovaries and Smad4flox/floxMerCreMer Stra8−/− ovaries

Project description:To confirm that female-to-male sexual fate reversal in Smad4flox/floxMerCreMer Stra8−/− ovaries occurs independently of somatic environment. We analyzed transcriptome of samples using RNA from control testes, ovaries, Smad4flox/floxMerCreMer Stra8+/− and Smad4flox/floxMerCreMer Stra8−/− ovaries.

2016-05-11 | GSE68773 | GEO

Transcriptional Metabolic Reprograming Drives Meiotic Fate Decision in Mammalian Germ Cells

Project description:Nitrogen starvation drives meiosis in yeasts1, while retinoic acid (RA) is required for mammalian meiosis through the activation of its germline-specific target, stimulated by retinoic acid gene 8 (Stra8).Here, by using single-cell transcriptome analysis of wild-type and Stra8-deficient testes, our data revealed that the expression of major metabolic genes are downregulated during germ cell entry into meiosis. Recently, we collected mouse testicular cells from wild-type and Stra8-deficient mice at postnatal day 12 (P12) and P21 for single cell RNA-sequencing (scRNA-seq). Upon scRNA-seq, we obtained a total of 23,470 cells from these samples. Using stringent quality control, 22,839 cells were selected for further analysis. All cells, including both wild-type and Stra8-deficient testicular cells, were pooled together to perform clustering analysis, which revealed 6 major cell clusters based on their distinct gene expression patterns. We annotated these six clusters by using known marker genes, including spermatogonia (SPG; Stra8; 3,941 cells), spermatocyte (SPC; Syce1; 4,924 cells), spermatid (STD; Acrv1; 710 cells), Sertoli cell (SC; Sox9; 9,488 cells), Myeloid (MYD; Acta2; 3,531 cells), and Leydig Cells (LY; Star; 245 cells). There is an increase in the percentage of spermatogonia in Stra8-deficient testes at both P12 (28% versus 14% in wild-type) and P21 (17% versus 10% in wild-type). At P21, while spermatocytes are abundant and spermatid are formed in wild-type testes, there are only a few spermatocytes (0.2% versus 73% in wild-type), and spermatids are absent in Stra8-deficient testes. Signature genes are enriched for GO terms for distinct biological function and metabolic pathways in different cell types.

2023-10-03 | GSE211390 | GEO

Characterization of molecular changes at meiotic initiation in mice

Project description:Characterization of the molecular changes occurring at murine meiotic initiation: identification of gene expression changes in male germ cells undergoing meiotic initiation; identification of STRA8-bound sites in mouse testes enriched for germ cells initiating meiosis

2019-02-26 | GSE115928 | GEO

Gene expression of mouse embryonic gonad [Nanos2_Stra8]

Project description:To examine whether the meiosis was solo factor responsible for the disruption of the male gonocyte differenitiation in the Nanos2-/- male germ cells, we deleted Stra8 function from Nanos2-null background and performed expression microarray analysis of the Nanos2+/-/Stra8+/- , Nanos2-/-/Stra8+/-, and Nanos2-/-/Stra8-/- male emnbryonic gonads at E14.5.

2013-02-06 | GSE38294 | GEO

Splicing factor SRSF1 is essential for homing of precursor spermatogonial stem cells in mice

Project description:Spermatogonial stem cells (SSCs) provide a continuous spermatogenesis and male fertility. However, the underlying mechanisms of alternative splicing (AS) in mouse SSCs are still largely unclear. We demonstrated that SRSF1 is essential for gene expression and splicing in mouse SSCs. Specific deletion of Srsf1 in mouse germ cells impairs self-renewal and homing of SSCs leading to male infertility. Whole-mount staining data showed the absence of germ cells in the testes of adult cKO mice, which indicates severe non-obstructive azoospermia (NOA) in cKO mice. Expression of SSC-related genes (Gfra1, Pou5f1, Plzf, Dnd1, Stra8, and Taf4b) was significantly reduced in the testes of conditional knockout (cKO) mice. CLIP-seq data found that SSC-related genes (Plzf, Id4, Setdb1, Stra8, Tial1, Bcas2, Ddx5, Srsf10, Uhrf1, and Bud31) were bound by SRSF1 in the mouse testes. Moreover, multi-omics analysis suggests that SRSF1 directly binds and regulates the expression of Tial1 via AS to implement SSCs self-renewal and differentiation. Collectively, our data reveal the critical role of an SRSF1-mediated AS in SSCs self-renewal and differentiation, which may provide a framework to elucidate the molecular mechanisms of the post-transcriptional network underlying SSCs.

2024-01-08 | GSE227575 | GEO

TDRD6 instructs correct assembly of mRNPs in the chromatoid body to potentiate UPF1 dependent mRNA decay.

Project description:RNA sequencing of heterozygote or Tudor domain contian protein 6 (TDRD6) knockout round spermatid cells. Chromatoid bodies (CBs) are germ cell-specific organelles of largely unknown function. CBs harbor various RNA species, RNA-associated proteins and proteins of the tudor domain family such as TDRD6. Proteome analysis of purified CBs revealed components of the nonsense-mediated mRNA decay machinery such as UPF1. TDRD6 is essential for UPF1 localization to CBs, for UPF1-UPF2 interaction, and for assembly of UPFs and other RNA binding proteins into super-complexes. In absence of TDRD6, the association of some mRNAs with UPF1 is impaired, and the long 3â UTR-stimulated but not the exon junction complex-stimulated pathway of NMD is distorted. Reduced association of mRNAs with UPF1 correlated with increased stability and presence in polysome fractions, i.e. enhanced translational activity. Thus, we define CBs as sites of UPF1-dependent mRNA degradation and provide evidence for the requirement for NMD in spermiogenesis. This function of CBs depends on TDRD6-promoted assembly of mRNA decay enzymes within mRNPs. RNA was extracted from quadruplicate samples and libraries generated for sequencing using the NEBNext Ultra Directional RNA Library Prep Kit (New England Biolabs) at the Deep Sequencing Group SFB 655, Biotechnology Center of Technische UniversitÃ¤t Dresden. After enrichment and XP bead (Agencourt AMPure Kit; Beckman Coulter, Inc.) purification, quality control was done using Fragment AnalyzerTM (Advanced Analytical). The bar-coded libraries were equimolarly pooled and subjected to 76 bp single-end sequencing on Illumina HiSeq 2000, resulting in an average of 33 million reads per sample.

2015-12-08 | E-GEOD-63948 | biostudies-arrayexpress

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