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Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse WT and Nxf2 KO post-natal day 21 testes

ABSTRACT: In euakryotes, mRNAs must be exported from the nucleus to the cytsoplasm. NXF2 is highly expressed in the mouse male germ cells. We are interested in its function in spermatogenesis, espically in the nuclear RNA export in the testis. To this end, we made Nxf2 mutant mice by gene targeting. In an attempt to identify the mRNA substrates of NXF2, we perform the microarray experiments on testes. We used microarrays to check the expression profiles of the Nxf2 WT and KO 21d testes on C57BL/6 background. Experiment Overall Design: To examine the expression difference between WT and Nxf2 KO testes, we collected testes from juvenile mice of three ages ( 21d, 26d, 28d). Testis weight was similar between WT and KO mice at post-natal day 21. Three pairs of WT and KO 21d testes were chosen for microarray analysis.

ORGANISM(S): Mus musculus

SUBMITTER: Jieyan Pan

PROVIDER: E-GEOD-13526 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcript profiling of WT and Nxf2 KO post-natal day 21 testes

Project description:In euakryotes, mRNAs must be exported from the nucleus to the cytsoplasm. NXF2 is highly expressed in the mouse male germ cells. We are interested in its function in spermatogenesis, espically in the nuclear RNA export in the testis. To this end, we made Nxf2 mutant mice by gene targeting. In an attempt to identify the mRNA substrates of NXF2, we perform the microarray experiments on testes. We used microarrays to check the expression profiles of the Nxf2 WT and KO 21d testes on C57BL/6 background.

2009-11-05 | GSE13526 | GEO

Upf2 in NMD pathway

Project description:Purpose: Probe the transcriptome-wide changes in the expression pattern between WT and Sertoli-specific Upf2 KO testes Methods: Total RNA were extracted from WT and Sertoli-specific Upf2 KO testes in triplicates and subject to deep-sequencing in Ion Torrent seq platform. Results: Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to the mouse genome (build mm9) and identified 16,014 transcripts in the retinas of WT and Nrl−/− mice with BWA workflow and 34,115 transcripts with TopHat workflow. RNA-seq data confirmed stable expression of 25 known housekeeping genes, and 12 of these were validated with qRT–PCR. RNA-seq data had a linear relationship with qRT–PCR for more than four orders of magnitude and a goodness of fit (R2) of 0.8798. Approximately 10% of the transcripts showed differential expression between the WT and Nrl−/− retina, with a fold change ≥1.5 and p value <0.05. Altered expression of 25 genes was confirmed with qRT–PCR, demonstrating the high degree of sensitivity of the RNA-seq method. Hierarchical clustering of differentially expressed genes uncovered several as yet uncharacterized genes that may contribute to retinal function. Data analysis with BWA and TopHat workflows revealed a significant overlap yet provided complementary insights in transcriptome profiling. Conclusions: Our study represents the first detailed analysis of Upf2-mediated NMD pathway in Sertoli cell development Testis mRNA profiling was generated from postnatal day 4 WT and Amh-cKO (Sertoli specific Upf2 KO) testes, in triplicates.

2014-09-06 | E-GEOD-58405 | biostudies-arrayexpress

Impaired spermatogenesis, muscle and erythrocyte function in U12 intron splicing-defective Zrsr1 mutant mice

Project description:We analysed the gene expression in testis of Zrsr1 mutant mice. We generated mutant mice containing truncating mutations within the RNA-recognition motif (RRM) in Zrsr1 locus. The RNAs of testes from 4 WT and 5 Zrsr1 mutant mice at day 15, immediately after the peak of expression of Zrsr1 in developing testis, were prepared and sequenced in order to identify and characterize the RNAs regulated by Zrsr1 in developing testis.

2018-04-03 | E-MTAB-4872 | biostudies-arrayexpress

Single-cell RNA-seq of total cells from testes of adult WT and miR-202-/- mice

Project description:We report to investigate the changes in cellular composition and gene expression upon miR-202 KO. One testis from each of three adult WT mice and three adult KO mice was collected, and the KO and WT testes were pooled respectively for subsequent analysis. A total of 15,310 WT and 12,935 KO cells passed standard quality control and were used for subsequent analysis.

2021-05-21 | GSE174731 | GEO

Testis-to-ovary reprogramming and testis regression after ablation of Sox9 in Sox8-/- mice

Project description:We used two Sertoli-cell-specific Cre lines: Wt1-CreERT2 and Sox9-CreERT2, to induce Sox9 ablation on a Sox8 -/- background in the adult testis. Sox9/8 double KO testes undergo testis-to-ovary genetic reprogramming and Sertoli-to-granulosa cell transdifferentiation.

2016-06-08 | E-MTAB-4572 | biostudies-arrayexpress

Taf7l cooperates with Trf2 to regulate spermiogenesis

Project description:Taf7l (a paralogue of Taf7) and Trf2 (a TBP-related protein) are components of the core promoter complex required for gene/tissue-specific transcription of protein-coding genes by RNA polymerase II. Previous studies reported that Taf7l knockout mice exhibit structurally abnormal sperm, reduced sperm count, weakened motility and compromised fertility. Here we find that continued backcrossing of Taf7l-/Y mice from N5 to N9 produced KO males that are essentially sterile. Genome-wide expression profiling by mRNA-seq analysis of wild type (WT) and Taf7l-/Y (KO) testes revealed that Taf7l ablation impairs the expression of many post-meiotic spermatogenic specific as well as metabolic genes. Importantly, histological analysis of testes revealed that Taf7l-/Y mice develop post-meiotic arrest at the first stage of spermiogenesis, phenotypically similar to Trf2-/- mice, but distinct from Taf4b-/- mice. Indeed, we find that Taf7l and Trf2 co-regulate post-meiotic genes, but none of Taf4b-regulated germ stem cell genes in testes. Genome-wide ChIP-seq studies indicate that TAF7L binds to promoters of activated post-meiotic genes in testis. Moreover, biochemical studies show that TAF7L associates with TRF2 both in vitro and in testis suggesting that TAF7L likely cooperates directly with TRF2 at promoters of a subset of post-meiotic genes to regulate spermiogenesis. Our findings thus provide a new mechanism for cell-type specific transcriptional control involving an interaction between a ‘non-prototypic’ core promoter recognition factor (Trf2) and an orphan TAF subunit (Taf7l) in mammalian testis-specific gene transcription. Genome-wide mapping of TAF7L and Pol II in testis tissue, and mRNA-seq expression profiling wild type and Taf7l knockout testis.

2013-09-29 | E-GEOD-50807 | biostudies-arrayexpress

Expression profiling of BORIS knockout mice testis

Project description:To identify differentially expressed genes in Boris KO testis, gene expression profilings were performed with NIAID mouse customized arrays. Gene expressions of five biological replicates of Boris KO testis were compared to those of wild type testis. Total RNAs from WT and BORIS KO testes prepared from P14 mice were labelled with Cy5 and Cy3, respectively. Five sets of each genotype were studied.

2010-06-05 | E-GEOD-19162 | biostudies-arrayexpress

PRM-based quantitative proteomic analysis of mouse tissues, testicular cells and cell lines

Project description:PRM analysis of RPL39L, RPL10L and RPL3L in 9 mouse tissues, including testis, fat, brain, liver, kidney, lung, skeletal muscle, heart and spleen; PRM analysis of RPL39 and RPL39L, RPL10 and RPL10L, and RPL22 and RPL22L1 in testicular cells; PRM analysis of RPL39L and RPL39 in RibosomeRPL39L-/- and wild type mouse spermatocytes and spermatids; PRM analysis of 17 down-regulated proteins and 1 unchanged protein (DYNC1LI2) in RibosomeRPL39L-/- testes; PRM analysis of RPL39 and RPL39L in GC-1 ribosomes, and RibosomeRPL39L-/- and wild type N2a ribosomes, Consistency between the observed and expected signal ratios of the light and heavy peptides by PRM in serial dilution experiments. Bovine serum albumin (BSA) as a negative control.

2022-09-30 | PXD020922 | Pride

Differential protein expression profiles of testes in Tmprss12-/- and wild-type mice

Project description:Serine protease is considered a relative upstream regulator in the physiological processes including male reproduction. Transmembrane serine protease 12 (TMPRSS12) has been shown to regulate sperm motility and uterotubal junction migration in mice, but its role in the testis remains unknown.In this study, we confirmed the important role of TMPRSS12 in spermatogenesis. To further investigate the mechanism of TMPRSS12 in male reproduction, we used 2-D electrophoresis combined with Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry(MALDI-TOF/TOF) to construct differential protein expression profiles of testes in Tmprss12-/- and WT mice. A total of 64 spots with 2-fold or more differential expression between two groups were identified using MALDI-TOF/TOF. As a result, 54 unique proteins were identified successfully, including 39 proteins with decreased expression and 15 proteins with increased expression in Tmprss12-/- mice.

2022-05-20 | PXD028014 | Pride

Identification of testis-relevant genes from testes ESTs and cDNA microarray in the black tiger shrimp

Project description:Identification of testis-relevant genes using in silico analysis from testes ESTs and cDNA microarray in the black tiger shrimp (Penaeus monodon) A total of 4,803 ESTs from the testis library were unidirectionally sequenced and analyzed in silico using a customizable data analysis package, ESTplus. A total of 2,702 unique sequences from 424 contigs and 2,278 singletons were identified. Approximately, 1,134 sequences from the total of 2,702 unique sequences (contigs and singletons) are homologous to genes with known functions. Some of which (mago nashi, insulin-degrading enzyme, actin-depolymerizing factor) are related to testicular development in studies of other organisms. Moreover, the sequences were further characterized according to the gene ontology categories (41% biological process, 24% molecular function, 35% cellular component). By comparing with the EST libraries of other tissues, a total of 1,579 possible testis-specific ESTs were identified. Some of these testis-specific ESTs, such as Phospholipase A2 and Matrix Metalloproteinases, have functions relevant to testicular development. In addition, novel ESTs (621 ESTs) that have never been identified in any ESTs from the Penaeidae family were found in this study. Through cDNA microarray analysis, some of testis-specific ESTs were preferentially expressed in testes to ovary; one of which was a saposin homolog. Therefore, saposin was further characterized by real-time PCR and its full-length sequence was obtained by RACE-PCR. A cDNA microarray was constructed from the EST libraries of P. monodon, consisting of 5,376 features (1,125 features were amplified from testis EST libraries). RNA samples were extracted from testes and ovaries of juveniles (4 month-old, pooled from n=112 and n=98 for testes and ovaries, respectively) and broodstocks from Andaman Sea. The RNA from ovary was labeled with Cy3 dye as a reference and that from testes was labeled with Cy5 dye.

2010-11-04 | E-GEOD-19037 | biostudies-arrayexpress

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