MicroRNA profiling in male rat germ cells after chronic cyclophosphamide treatment
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ABSTRACT:
PROVIDER: PRJNA489300 | ENA |
REPOSITORIES: ENA
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Project description:Although numerous miRNAs have been identified in the testis, their roles in regulating the highly specific events that occur in the different germ cell types throughout spermatogenesis remain largely unknown. Furthermore, whether male germ cell miRNA expression is altered in response to or as a consequence of exposure to a toxic agent is unknown. Here we examine miRNA expression profiles in pachytene spermatocytes and round spermatids obtained from control rats and from rats treated with a chronic low dose of cyclophosphamide, a male germ cell toxicant. We observed that pachytene spermatocytes and round spermatids display vastly different miRNA expression profiles, reflecting their different developmental stages and possibly influencing the cellular response to toxic insult. Chronic low dose cyclophosphamide treatment altered the miRNA profiles in both pachytene spermatocytes and round spermatids. Target prediction analyses revealed that miRNAs altered by cyclophosphamide treatment may be involved in the response to cellular stress and damage. However, many are also involved in processes that are crucial for proper germ cell development. This study suggests that pachytene spermatocytes and round spermatids display distinct miRNA profiles that can be altered by cyclophosphamide treatment. The observed changes may be part of a response and repair mechanism to cyclophosphamide-induced damage or a dysregulation that disrupts normal germ cell development.
2018-09-05 | GSE119473 | GEO
Project description:To further understand the effects of chronic cyclophosphamide treatment on spermatogenesis, we used whole genome microarrays to identify differentially expressed genes in pachytene spermatocytes and round spermatids from treated and control male rats.
2016-03-23 | GSE79471 | GEO
Project description:To further understand the effects of chronic cyclophosphamide treatment on spermatogenesis, we used whole genome microarrays to identify differentially expressed genes in pachytene spermatocytes and round spermatids from treated and control male rats. Pachytene spermatocytes and round spermatids from rats treated chronically with cyclophosphamide were isolated and profiled for changes in gene expression.
2016-03-23 | E-GEOD-79471 | biostudies-arrayexpress
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2023-06-19 | GSE220440 | GEO
Project description:Spermatogenesis is a complex process, dependent upon the successive activation and/or repression of thousands of gene products, and ends with the production of haploid male gametes. RNA sequencing of male germ cells in the rat identified thousands of novel testicular unannotated transcripts, named TUTs. Although such RNAs are usually annotated as long non-coding RNAs, it is possible that some of these TUTs code for protein. To test this possibility, we used a “Proteomics Informed by Transcriptomics” strategy, combining shotgun proteomics analyses and RNA sequencing data for enriched populations of rat testicular cells. Among 3559 TUTs and 506 lncRNAs found in meiotic and post-meiotic germ cells, 44 encoded at least one peptide. We show that these novel high-confidence protein-coding loci exhibit several genomic features intermediate between those of lncRNAs and mRNAs. We experimentally validated the testicular expression pattern of two of these novel protein-coding gene candidates, both highly conserved in mammals: one for a vesicle-associated membrane protein, we named VAMP-9, and the other for an enolase domain-containing protein. This study confirms the potential of PIT approaches for the discovery of protein coding transcripts initially thought to be untranslated, or unknown transcripts. Our results contribute to the understanding of spermatogenesis by characterizing two novel proteins, implicated by their strong expression in germ cells.
2014-09-12 | PXD000872 | Pride
Project description:DNA methylation dynamic in male rat germ cells during gametogenesis
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Project description:Male germ cell meiosis is essential for generating haploid spermatozoa in mice. Here, we investigate the essential role of DIS3L2 in male germ cell meiosis in mice. Conditional inactivation of DIS3L2 in spermatocytes with Stra8-cre transgenic mice have severely impaired meiotic progression, which results in defective meosis and spermatogenesis associated with a Sertoli cell-only syndrome and adult sterility. RNA-seq analysis reveales that Dis3l2 deficiency causes significant dysregulation of the expression of transcripts in mutant testes. Meiosis-assocaited genes are significantly decreased in the absence of DIS3L2. Therefore, we show that DIS3L2 ribonuclease plays a critical role in germ cell meiosis during spermatogenesis in mice.
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Project description:Heterogeneity within murine prospermatogonia manifests as certain populations either die or survive during a developmentally scheduled apoptotic event at E13.5. We have identified that this apoptosis is clonal, suggesting that apoptotic fate is heritably enforced among populations of germ cells. To determine how subpopulations of germ cells differ, we used single-cell RNA sequencing to identify two diametric states: an apoptotic (AP) versus a male-differentiated (MD) subpopulation that differ primarily by the expression of critical male-differentiation genes known to be epigenetically regulated. We isolated AP and MD germ cells from E13.5 male embryos for whole genome bisulfite sequencing to compare how methylation differs between these populations. We find that AP germ cells are relatively hypermethylated at a whole-genome level and, importantly, are significantly hypermethylated at specific genome-reprogramming responsive (GRR) loci. Because these GRR genes facilitate male differentiation, we conclude that inappropriate demethylation as germ cells epigentically reprogram in the period preceding apoptosis produces an aberrantly male-differentiated clone that is fated for elimination. This quality-control paradigm therefore links fidelitous epigenetic reprogramming to germ cell selection.
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