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Functional diversity and convergence of small non-coding RNAs in male germ cell differentiation and fertilization (piRNA and endo-siRNAs)

ABSTRACT: The small non-coding RNAs (sncRNAs) are considered as postranscriptional key regulators of male germ cell development. In addition to microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), other sncRNAs generated from small nucleolar RNAs (snoRNAs), tRNAs or rRNAs processing may also play important regulatory roles in spermatogenesis. By next generation sequencing (NGS), we characterized the different sncRNA populations detected at three milestone stages in male germ differentiation: primordial germ cells (PGCs) at 13.5 dpc, pubertal spermatogonia cells, and mature spermatozoa. In order to assess the potential transmission of the sncRNAs through the mature spermatozoa during fertilization, the sncRNA population detected in male germ cells was also compared with sncRNAs detected in unfertilized mouse oocytes and zygotes. Combining the data obtained by NGS and microarrays from whole PGC and spermatogonia transcriptome, we defined the potential regulatory roles of specific miRNAs and their validated targets. Similar to miRNAs, both the small RNAs derived from snoRNAs and the piRNAs, could be involved in the postranscriptional regulation of mRNA transcripts during the male germ development. Finally, our results strongly suggest that the small RNAs-derived from tRNAs and rRNAs are interacting with PIWI proteins, and specifically with MILI. These new classes of piRNAs are not generated by the ping-pong pathway and could be the source of primary piRNAs.

ORGANISM(S): Mus musculus

PROVIDER: GSE59254 | GEO | 2015/03/26

SECONDARY ACCESSION(S): PRJNA254814

REPOSITORIES: GEO

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Functional diversity and convergence of small non-coding RNAs in male germ cell differentiation and fertilization (piRNA and endo-siRNAs)

Project description:The small non-coding RNAs (sncRNAs) are considered as postranscriptional key regulators of male germ cell development. In addition to microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), other sncRNAs generated from small nucleolar RNAs (snoRNAs), tRNAs or rRNAs processing may also play important regulatory roles in spermatogenesis. By next generation sequencing (NGS), we characterized the different sncRNA populations detected at three milestone stages in male germ differentiation: primordial germ cells (PGCs) at 13.5 dpc, pubertal spermatogonia cells, and mature spermatozoa. In order to assess the potential transmission of the sncRNAs through the mature spermatozoa during fertilization, the sncRNA population detected in male germ cells was also compared with sncRNAs detected in unfertilized mouse oocytes and zygotes. Combining the data obtained by NGS and microarrays from whole PGC and spermatogonia transcriptome, we defined the potential regulatory roles of specific miRNAs and their validated targets. Similar to miRNAs, both the small RNAs derived from snoRNAs and the piRNAs, could be involved in the postranscriptional regulation of mRNA transcripts during the male germ development. Finally, our results strongly suggest that the small RNAs-derived from tRNAs and rRNAs are interacting with PIWI proteins, and specifically with MILI. These new classes of piRNAs are not generated by the ping-pong pathway and could be the source of primary piRNAs. Comparative analysis from deep sequencing of piRNAs and endo-siRNAs in mouse oocytes, spermatozoa and zygotes

2015-03-26 | E-GEOD-59254 | biostudies-arrayexpress

Functional diversity and convergence of small non-coding RNAs in male germ cell differentiation and fertilization (mRNA analysis)

Project description:The small non-coding RNAs (sncRNAs) are considered as postranscriptional key regulators of male germ cell development. In addition to microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), other sncRNAs generated from small nucleolar RNAs (snoRNAs), tRNAs or rRNAs processing may also play important regulatory roles in spermatogenesis. By next generation sequencing (NGS), we characterized the different sncRNA populations detected at three milestone stages in male germ differentiation: primordial germ cells (PGCs) at 13.5 dpc, pubertal spermatogonia cells, and mature spermatozoa. In order to assess the potential transmission of the sncRNAs through the mature spermatozoa during fertilization, the sncRNA population detected in male germ cells was also compared with sncRNAs detected in unfertilized mouse oocytes and zygotes. Combining the data obtained by NGS and microarrays from whole PGC and spermatogonia transcriptome, we defined the potential regulatory roles of specific miRNAs and their validated targets. Similar to miRNAs, both the small RNAs derived from snoRNAs and the piRNAs, could be involved in the postranscriptional regulation of mRNA transcripts during the male germ development. Finally, our results strongly suggest that the small RNAs-derived from tRNAs and rRNAs are interacting with PIWI proteins, and specifically with MILI. These new classes of piRNAs are not generated by the ping-pong pathway and could be the source of primary piRNAs. mRNA analysis of Primordial Germ Cells (PGCs), Spermatogonia cells (SPG), adult testis (AdT) and Gonad-less (GL) embryos. Indirect comparisons were made across multiple arrays with raw data pulled from different channels for data analysis and comparison to the control data.

2015-03-26 | E-GEOD-59252 | biostudies-arrayexpress

Functional diversity and convergence of small non-coding RNAs in male germ cell differentiation and fertilization (mRNA analysis)

2015-03-26 | GSE59252 | GEO

Small non-coding RNA discovery and profiling with sRNAtools based on high-throughput sequencing

Project description:Small non-coding RNAs (sncRNAs) are generated from different genomic loci and have important roles in biological processes, such as cell proliferation and regulation of gene expression. Next-generation sequencing (NGS) has provided an unprecedented opportunity to discover and quantify diverse kinds of sncRNAs, such as tRFs (tRNA-derived small RNA fragments), phasiRNAs (phased, secondary, small interfering RNAs), piRNAs and plant specific 24-nt short interfering RNAs (siRNAs). However, currently available web-based tools do not provide approaches to comprehensively analyze all these diverse sncRNAs. A novel web server, sRNAtools (https://bioinformatics.sc.cn/sRNAtools), is presented that can be used in conjunction with high-throughput sequencing to discover, profile and functionally annotate sncRNAs, including miRNAs, isomiRs, piRNAs, tRFs, tRNAs, snRNAs, snoRNAs, rRNAs, phasiRNAs, and plant specific 24-nt siRNAs for 21 species. In this study, sRNAtools has been used in sncRNA profiling in poplar wood tissues, which identified many wood related sncRNAs expression patterns.We believe that sRNAtools will greatly benefit the scientific community as an integrated tool to study sncRNAs.

2019-11-15 | GSE139897 | GEO

Profiling the Small Non-coding RNA Transcriptome of the Human Placenta

Project description:Proper functioning of the placenta, the temporary organ crucial in fetal development during pregnancy, is critical for maternal and fetal health. While microRNAs (miRNAs) are known to impact placental gene expression, the effects of other small non-coding RNAs (sncRNAs) on the placental transcriptome are not well-established. Here, we performed small RNA sequencing of 30 chorionic villi samples of varying gestational ages (M = 23.5 ± 11.4 weeks). We observed expression of 1543 sncRNAs, including miRNAs, piRNAs, snRNAs, snoRNAs, and tRNAs, as well as 18,003 miRNA variants (isomiRs), and identified 48 previously-unannotated miRNAs. This novel characterization of the placental sncRNA transcriptome may aid future investigations of sncRNA regulatory functions within the human placenta.

2021-05-14 | GSE164178 | GEO

Unraveling mitochondrial piRNAs in mouse embryonic gonadal cells

Project description:Although mitochondria are widely studied organelles, the recent interest in the role of mitochondrial small non-coding RNAs (sncRNA) is providing new functional perspectives in germ cell development and differentiation. PIWI-interacting RNAs (piRNAs) are single-stranded sncRNAs of 20-35 nt generated from the processing of pre-piRNAs longer molecules to be functionally bound to PIWI proteins. Initially ascribed to germ cells, as protectors against transposons, we now know that they are also expressed in somatic cells. In mammals, germ cells differentiate at early stages of embryonic development in the primitive gonads as primordial germ cells (PGCs), initiating divergent pathways in both sexes, accompanied by somatic cells (SCs) of the ovary or testis. Previously, we identified mitochondrial piRNAs (mito-piRNAs) in mouse germ and somatic cells. However, a comparative analysis of mito-piRNAs between PGCs and SCs, between sexes and during early gonadal development has not been performed. We leverage NGS data obtained from PGCs and SCs purified from early differentiating embryonic ovaries and testis from E11.5 to E13.5. Using bioinformatic tools, here we unravel: the origin (from nuclear or mitochondrial genome), the levels of expression, the potential role of mito-piRNAs, as well as their association with genomic regions encoding other sncRNAs (such as tRNAs and rRNAs) and the mitochondrial regulatory region (D-loop). Finally, our results indicate nucleo-mitochondrial communication at both anterograde and retrograde signaling, mediated by mito-piRNAs.

2021-07-01 | GSE179299 | GEO

P6 Mouse Sertoli Cell Small RNA seq

Project description:This study annotates the NGS reads in a Sertoli cell sncRNA library. In addition to known sncRNAs, we also identified numerous novel sncRNAs expressed by Sertoli cells. Our data suggest that the Sertoli cell sncRNA transcriptome predominantly consists of miRNAs, piRNA-like RNAs, tRNAs and snoRNAs. This study reports the first comprehensive annotation of the Sertoli cell sncRNA transcriptome. Sertoli cells were purified from post-natal day 6 C57Bl/6 mice using the STAPUT unit gravity sedimentation apparatus. Small RNA was isolated from cells and a cDNA library was generated for 454 sequencing. Sequences from FastA files were initially mapped to known sequences and the genome and subsequently annotated upon novel small RNA categorization.

2012-09-08 | E-GEOD-40692 | biostudies-arrayexpress

P6 Mouse Sertoli Cell Small RNA seq

2012-09-08 | GSE40692 | GEO

Small RNA profiling reveals deregulated PTEN/PI3K/Akt pathway in asthmatic bronchial smooth muscle cells

Project description:Background: Aberrant expression of small non-coding RNAs (sncRNAs), in particular microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs) define several pathological processes. Asthma is characterized by airway hyper-reactivity, chronic inflammation and airway wall remodeling. Asthma-specific miRNA profiles were reported for bronchial epithelial cells, but no information on sncRNA expression in asthmatic bronchial smooth muscle (BSM) cells is available. Objective: To determine whether primary BSM sncRNA expression profile is altered in asthma and identify targets of differentially expressed sncRNAs. Methods: SmallRNA sequencing was used for sncRNA profiling in BSM cells (8 asthma, 6 non-asthma). sncRNA identification and differential expression analysis was performed with iMir, . experimentally validated miRNA targets were identified with Ingenuity Pathway Analysis and putative piRNA targets with miRanda. Results: Asthmatic BSM cells showed abnormal expression of 32 sncRNAs (26 miRNAs, 5 piRNAs, and 1 snoRNA). Target prediction for deregulated miRNAs and piRNAs revealed experimentally validated and predicted mRNA targets expressed in the BSM cells. 38 of these mRNAs represent major targets for deregulated miRNAs and may play important roles in the pathophysiology of asthma. Interestingly, 6 such miRNAs were previously associated with asthma and/or considered as novel therapeutic targets for treatment of this disease. Signaling pathway analysis revealed involvement of these sncRNAs in increased cell proliferation via PTEN and PI3K/Akt signaling pathways. Conclusions: BSM cells from asthma patients are characterized by aberrant sncRNA expression that recapitulates multiple pathological phenotypes of these cells. Implications: sncRNA expression profiling performed in this study further improve our understanding of the molecular mechanisms underlying asthma-associated processes in lungs.

2018-01-07 | GSE64744 | GEO

Characterization of the small RNA population in Human spermatozoa

Project description:Small noncoding RNAs (sncRNA) are becoming recognized for their participation in a diverse range of cellular functions. In this study, their global characterization in human spermatozoa from donors with proven fertility was undertaken. Reads were analyzed in two classes, those mapping to unique locations and those that could be aligned to up to 10 genomic locations. All libraries showed comparable distribution of reads between intergenic, intronic and exonic genomic regions. Analysis of the sequences revealed the presence of multiple small RNA classes. The miRNAs retained in spermatozoa were found to be associated with promoter regions, suggestive of a role at the transcriptional level. Piwi-interacting sncRNAs as well as repeat-associated small RNAs were identified for the first time in mature spermatozoa from a mammalian species. Human spermatozoa retain a multifaceted population of small non-coding RNAs. Their presence is consistent with the view that they may function to stabilize the genome as part of the confrontation and consolidation of the genomes at fertilization. Examination of sperm samples from 3 individuals, sequenced individually

2011-10-15 | E-GEOD-21191 | biostudies-arrayexpress

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