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The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis [mRNAseq]

ABSTRACT: The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis [mRNAseq]

PROVIDER: PRJNA922909 | ENA |

REPOSITORIES: ENA

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The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis [mRNAseq]

Project description:GRTH/DDX 25 is a member of the DEAD-box family of RNA helicases that play an essential role in spermatogenesis. Regulation of spermatogenesis occurs at the levels of transcription, post-transcriptional and translational levels which needs to be explored in detail. miRNAs regulate the expression of important genes involved in spermatid elongation process. Differential miRNA expression in round spermatids (RS) of GRTH KO and GRTH-KI mice in comparison with mRNAs remains unexplored. Our miRNAseq analysis of small RNAs obtained from RS reveal differential expression of important miRNAs that regulate spermatid differentiation, apoptosis, chromatin compaction and ubiquitination. mRNAseq analysis of RS isolated from GRTH-KI and GRTH-KO mice models were also carried-out to study the putative miRNA targets and mRNA-miRNA interaction. mRNAseq analysis revealed specific set of mRNAs that are critical for sperm chromatin compaction, ubiquitination and spermatid elongation etc. miRNA-mRNA interaction prediction highlighted differentially expressed/enriched miRNA transcripts whose predicted mRNA targets were differentially expressed. Based on the interaction information of the miRNAs-mRNAs differential analysis, the miRNA-mRNA network was constructed. These results demonstrate the importance of miRNA in the translational arrest and stability of germ cell specific mRNAs which are critical for later stages of spermiogenesis and fertility.

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The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis [miRNAseq]

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The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis

Project description:The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis

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The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis [miRNAseq]

Project description:The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis [miRNAseq]

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miRNA Expression Profiles of Mouse Round Spermatids in GRTH/DDX25-Mediated Spermiogenesis: mRNA-miRNA Network Analysis.

Project description:GRTH/DDX25 is a testis-specific DEAD-box family of RNA helicase, which plays an essential role in spermatogenesis and male fertility. There are two forms of GRTH, a 56 kDa non-phosphorylated form and a 61 kDa phosphorylated form (pGRTH). GRTH-KO and GRTH Knock-In (KI) mice with R242H mutation (lack pGRTH) are sterile with a spermatogenic arrest at step 8 of spermiogenesis due to failure of round spermatids (RS) to elongate. We performed mRNA-seq and miRNA-seq analysis on RS of WT, KI, and KO to identify crucial microRNAs (miRNAs) and mRNAs during RS development by establishing a miRNA-mRNA network. We identified increased levels of miRNAs such as miR146, miR122a, miR26a, miR27a, miR150, miR196a, and miR328 that are relevant to spermatogenesis. mRNA-miRNA target analysis on these DE-miRNAs and DE-mRNAs revealed miRNA target genes involved in ubiquitination process (Ube2k, Rnf138, Spata3), RS differentiation, and chromatin remodeling/compaction (Tnp1/2, Prm1/2/3, Tssk3/6), reversible protein phosphorylation (Pim1, Hipk1, Csnk1g2, Prkcq, Ppp2r5a), and acrosome stability (Pdzd8). Post-transcriptional and translational regulation of some of these germ-cell-specific mRNAs by miRNA-regulated translation arrest and/or decay may lead to spermatogenic arrest in KO and KI mice. Our studies demonstrate the importance of pGRTH in the chromatin compaction and remodeling process, which mediates the differentiation of RS into elongated spermatids through miRNA-mRNA interactions.

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Differential miroRNA expression in round spermatids of GRTH null mice compared to wild type.

Project description:miRNA expression profiles for round spermatids of wild type and GRTH knock-out mice were determined by Rodent TaqMan® Low Density miRNA Arrays A v2.0 (TLDA, Applied Biosystem). Purified round spermatids were prepared from the testis of wild type and GRTH null mice (C57BL/6 strain). Equal amount of total RNA from 20 mice (wild type or GRTH KO) was pooled prior to gene expression analysis.

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Linking Phospho-Gonadotropin Regulated Testicular RNA Helicase (GRTH/DDX25) to Histone Ubiquitination and Acetylation Essential for Spermatid Development During Spermiogenesis

Project description:In this study, we examined genome-wide transcriptome profiles of germ cells isolated from GRTH Knock-In and WT mice testis using RNA-Seq. We used homozygous KI mice with R242H mutation in GRTH gene resulting loss of phospho-GRTH to further understand its role in spermatid development during spermiogenesis by analyzing the DEGs from RNA-Seq. The analysis of RNA-Seq data allowed us to find the relevant DEGs related to ubiquitination pathway that are essential for spermatid development. This study using KI mice model demonstrated that loss of phospho-GRTH impairs UBE2J1 and RNF8 dependent histone modifications hampering initial stages of spermatid elongation process.

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Differential miroRNA expression in round spermatids of GRTH null mice compared to wild type.

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Role of phosphorylated Gonadotropin-Regulated Testicular RNA Helicase (GRTH/DDX25) in the Regulation of Germ Cell Specific mRNAs in Chromatoid Bodies during Spermatogenesis

Project description:In this study, we examined transcriptome profiles of Chromatoid bodies (organelle; CB) of germ cells isolated from GRTH Knock-In (Human GRTH gene with R242H mutation) and WT mice testis using RNA-Seq. We used homozygous GRTH KI mice (which show loss of phospho-GRTH) to further understand the importance of CB and phospho-GRTH and their role in spermatid development during spermiogenesis by analyzing the DEGs from RNA-Seq. The analysis of RNA-Seq data allowed us to find the relevant DEGs related to RNA Transport and Protein processing that are essential for spermatid development. This study using KI mice model demonstrated that loss of phospho-GRTH impairs CB structure and mRNA storage function (in CB) and chromatin compaction hampering initial stages of spermatid elongation process.

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The miRNA expression profiles of round spermatids during GRTH/DDX25 mediated spermiogenesis: mRNA-miRNA Network analysis

Project description:This SuperSeries is composed of the SubSeries listed below.

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