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Deep sequencing of small RNA facilitates tissue and sex associated microRNA discovery in zebrafish

ABSTRACT: The role of microRNAs in gene regulation has been well established. The extent of miRNA regulation also increases with increasing genome complexity. Though the number of genes appear to be equal between human and zebrafish, substantially less microRNAs have been discovered in zebrafish compared to human (Release 19). It appears that most of the miRNAs in zebrafish are yet to be discovered. We sequenced small RNAs from brain, gut, liver, ovary, testis, eye, heart and embryo of zebrafish. In brain, gut and liver sequencing was done in male and female separately. Majority of the sequenced reads (16-62%) mapped to known miRNAs, with the exception of ovary (5.7%) and testis (7.8%). Using the miRNA discovery tool (miRDeep2), we discovered novel miRNAs from the un-annotated reads that ranged from 7.6 to 23.0%, with exceptions of ovary (51.4%) and testis (55.2%). The prediction tool identified a total of 459 novel pre-miRNAs. We compared expression of miRNAs between different tissues and between males and females to identify tissue associated and sex associated miRNAs respectively. These miRNAs could serve as putative biomarkers for these tissues. The brain and liver had highest number of tissue associated (22) and sex associated (34) miRNAs, respectively. This study comprehensively identifies tissue and sex associated miRNAs in zebrafish. Further, we have discovered 459 novel pre-miRNAs (~30% seed homology to human miRNA) as a genomic resource which can facilitate further investigations to understand miRNA-mRNA gene regulatory networks in zebrafish which will have implications in understanding the function of human homologs.

ORGANISM(S): Danio rerio

PROVIDER: GSE57169 | GEO | 2015/10/16

SECONDARY ACCESSION(S): PRJNA245824

REPOSITORIES: GEO

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Deep sequencing of small RNA facilitates tissue and sex associated microRNA discovery in zebrafish

Project description:The role of microRNAs in gene regulation has been well established. The extent of miRNA regulation also increases with increasing genome complexity. Though the number of genes appear to be equal between human and zebrafish, substantially less microRNAs have been discovered in zebrafish compared to human (Release 19). It appears that most of the miRNAs in zebrafish are yet to be discovered. We sequenced small RNAs from brain, gut, liver, ovary, testis, eye, heart and embryo of zebrafish. In brain, gut and liver sequencing was done in male and female separately. Majority of the sequenced reads (16-62%) mapped to known miRNAs, with the exception of ovary (5.7%) and testis (7.8%). Using the miRNA discovery tool (miRDeep2), we discovered novel miRNAs from the un-annotated reads that ranged from 7.6 to 23.0%, with exceptions of ovary (51.4%) and testis (55.2%). The prediction tool identified a total of 459 novel pre-miRNAs. We compared expression of miRNAs between different tissues and between males and females to identify tissue associated and sex associated miRNAs respectively. These miRNAs could serve as putative biomarkers for these tissues. The brain and liver had highest number of tissue associated (22) and sex associated (34) miRNAs, respectively. This study comprehensively identifies tissue and sex associated miRNAs in zebrafish. Further, we have discovered 459 novel pre-miRNAs (~30% seed homology to human miRNA) as a genomic resource which can facilitate further investigations to understand miRNA-mRNA gene regulatory networks in zebrafish which will have implications in understanding the function of human homologs. Known miRNA profiling, novel miRNA discovery and identification of tissue associated and sex associated miRNAs from sRNA deep sequencing data of different tissues and embryo of zebrafish (in triplicate) was carried out using the Illumina HiSeq 2000 platform.

2015-10-16 | E-GEOD-57169 | biostudies-arrayexpress

sRNA sequencing of mature and developing gonads during protandrous changes in the Asian seabass

Project description:Purpose: sRNA-sequencing of mature and intermediate gonadal tissue in order to identify the differential expression of miRNAs during male to female (i.e. protandrous) sex transition Methods: Total RNA was extracted and sRNA was purified. cDNA libraries were constructed using a high definition adapter protocol (Xu et al. 2015). 50 bp sequencing was performed on Illumina's HiSeq 2500 at the Earlham Institute, Norwich, UK. Sequenced data was trimmed for adapters and filtered to remove very short and low complexity sequences. miRBase animal miRNA precursor sequences were mapped against the Asian seabass genome in order to generate a set of putative miRNA precursors. Putative precursor molecules with aligning mature miRBase miRNA(s) and forming a valid pre-miRNA hairpin structure were annotated as valid precursor miRNAs. Novel precursor and mature miRNAs were annotated using a combination of published algorithms and manual checking to ensure consistency with canonical miRNA biogenesis criteria. The alignment of sequenced reads against a non-redundant miRNA precursor set was used to determine raw read counts of mature miRNAs. Differential expression analysis was performed in order to identify differentially expressed mature miRNAs between conditions. Results: We detect 156, 71, 122, 151, 171 and 155 differentially expressed miRNA for the testis -> T1/T2, T1/T2 -> T3/T4, T3/T4 -> ovary, testis -> T3/T4, T1/T2 -> ovary and the testis->ovary comparisons respectively Conclusions: There is substantial differential expression of miRNAs at every stage of gonadal change in the Asian seabass during the sex transition process

2022-10-20 | GSE122720 | GEO

Identification of sex-biased miRNA involved in sex differentiation and spermatogenesis in yellow catfish by comparative transcriptome analysis

Project description:In this study, three small RNA libraries constructed from gonad tissues of XX female, XY male and YY super-male yellow catfish were sequenced by Solexa high-throughput sequencing technology to investigate the expression pattern of sex-biased microRNA. The sequencing data generated a total of 384 conserved miRNAs and 113 potential novel miRNAs, among which 23, 30 and 14 miRNAs were specifically detected in XX ovary, XY testis, and YY testis, respectively. Interestingly, more abundant piRNAs were found in ovary compared to testis in yellow catfish, which phenomenon is also observed in other fish species but opposite in mammalians. We detected a number of microRNAs differentially expressed between ovary and testis, such as miR-21, miR-462, miR-430 and -200 family. When compared the transcriptome between XY and YY testis, we observed relative lower expression of miR-141 and miR-429 in YY testis. Histological analysis indicated that YY super-males have more spermatids and less spermatocytes in spermatogenic cyst than XY males under the same age and culturing conditions. The expression level of miR-141 and 429 coincides with the progression of spermatogenesis both in yellow catfish and human. At last, The expression pattern of nine arbitrarily selected miRNAs detected by quantitative RT-PCR was consistent with the Solexa sequencing results. Our study provides a comprehensive miRNA transcriptome analysis for gonad of yellow catfish with different sex genotypes, and identifies a number of sex-biased miRNAs that are potentially involved in sex differentiation and spermatogenesis.

2016-01-24 | GSE54610 | GEO

Sex-biased miRNA expression in mouse, opossum and chicken

Project description:Sexual dimorphism depends on sex-biased gene expression, but the contributions of microRNAs (miRNAs) have not been globally assessed. We therefore produced an extensive small RNA sequencing dataset to analyse male and female miRNA expression profiles in mouse, opossum and chicken. Our analyses uncovered numerous cases of somatic sex-biased miRNA expression, especially in the mouse heart and liver. Sex-biased expression is explained by miRNA-specific regulation, including sex-biased chromatin accessibility at promoters, rather than piggybacking of intronic miRNAs on sex-biased protein-coding genes. In mouse, but not opossum and chicken, sex bias is coordinated across tissues such that autosomal testis-biased miRNAs tend to be somatically male-biased, whereas autosomal ovary-biased miRNAs are female-biased, possibly due to broad hormonal control. In chicken, which has a Z/W sex chromosome system, expression output of genes on the Z chromosome is expected to be male-biased, since there is no global dosage compensation mechanism that restores expression in ZW females after almost all genes on the W chromosome decayed. Nevertheless, we found that the dominant liver miRNA, miR-122-5p, is Z-linked but expressed in an unbiased manner, due to the unusual retention of a W-linked copy. Another Z-linked miRNA, the male-biased miR-2954-3p, shows conserved preference for dosage-sensitive genes on the Z chromosome, based on computational and experimental data from chicken and zebra finch, and acts to equalise male-to-female expression ratios of its targets. Unexpectedly, our findings thus establish miRNA regulation as a novel gene-specific dosage compensation mechanism.

2017-11-01 | GSE102062 | GEO

Repertoire of porcine microRNAs in adult ovary and testis by deep sequencing.

Project description:Despite increasing efforts in miRNAs identification across various species and diverse tissue types, little is known about porcine gonad-specific miRNAs. Although the well-known importance of pig in agriculture, as well as a model for human biology, the miRNA catalog of pig has been largely undefined. Identification and preliminary characterization of gonad-specific miRNAs would be a prerequisite for a thorough understanding of their roles in regulating folliculogenesis and spermatogenesis. In the present study, we get insight into miRNA transcriptome in adult porcine ovary and testis using deep sequencing technology, and to elucidate their characteristic organ- and gender-specific profiles, genomic context and emphasize the features of X-linked miRNAs. Two small RNA libraries from adult porcine ovary and testis tissues were sequenced.

2011-03-29 | E-GEOD-24443 | biostudies-arrayexpress

Identification of novel ovarian predominant miRNAs in medaka

Project description:The involvement of miRNAs during vertebrate oogenesis is poorly documented. Based on the assumption that ovarian-specific or ovarian-predominant genes usually play important roles during oogenesis, we searched for ovarian-predominant miRNAs in the medaka (Oryzias latipes) ovary. 10 tissus were collected from adult medaka (intestine, ovary, testis, liver, heart, gills, kidney, brain, muscle and bone) and RNAs were hybridized on a designed agilent microarray displaying 3800 distinct miRNAs from different teleost and vertebrate species. We identified 66 miRNAs sequences predominantly expressed in the ovary that had never been previously described in medaka.

2016-08-31 | GSE80823 | GEO

Regulation of the gonadal transcriptome during sex determination and testis morphogenesis: comparative candidate genes

Project description:Embryonic day 13 (E13), E14, and E16 rat testes and ovaries were used for microarray analysis, as well as E13 testis organ cultures that undergo testis morphogenesis and develop seminiferous cords in vitro. A list of 109 genes resulted from a selective analysis for genes present in male gonadal development and with a 1.5-fold change in expression between E13 and E16. Characterization of these 109 genes potentially important for testis development revealed that cytoskeletal-associated proteins, extracellular matrix factors, and signaling factors were highly represented. Throughout the developmental period (E13-E16), sex-enriched transcripts were more prevalent in the male with 34 of the 109 genes having testis-enriched expression during sex determination. In ovaries, the total number of transcripts with a 1.5-fold change in expression between E13 and E16 was similar to the testis, but none of those genes were both ovary enriched and regulated during the developmental period. Genes conserved in sex determination were identified by comparing changing transcripts in the rat analysis herein, to transcripts altered in previously published mouse studies of gonadal sex determination. A comparison of changing mouse and rat transcripts identified 43 genes with species conservation in sex determination and testis development. Profiles of gene expression during E13-E16 rat testis and ovary development are presented and candidate genes for involvement in sex determination and testis differentiation are identified. Analysis of cellular pathways did not reveal any specific pathways involving multiple candidate genes. However, the genes and gene network identified influence numerous cellular processes with cellular differentiation, proliferation, focal contact, RNA localization, and development being predominant. Keywords: expression analysis, testis, ovary, sex determination

2008-02-20 | GSE10558 | GEO

Annotation of the zebrafish genome through an integrated transcriptomic and proteomic analysis

Project description:Zebrafish organ proteomics was carried out for proteogenomic analysis. High resolution mass spectrometry-based proteomic profiling of 11 adult organs (eye, brain, liver, spleen, intestine-pancreas, ovary, testes, muscle, heart and head) and two developmental stages (embryos 48 and 120 hours post-fertilization) of zebrafish (SAT line) was carried out. Protein extracts were reduced and alkylated (iodoacetamide). Off-loine fractionation was carried out by SDS-PAGE and basic RPLC. MS analysis was cariied on Agilent 6540 Q-TOF. Spleen and Testis samples were also analyzed on Thermo Orbitrap LTQ Velos.

2014-02-26 | PXD000479 | Pride

Sex Specific Transciption in Mouse Tissues

Project description:Sex Specific Transciption in Mouse Hypothalamus, Liver, Kidney, Ovary and Testis. For each somatic tissue there are 3 biological samples from different pools comprised of 10 animals for each sex each with a technical replicate. For each of the reproductive tissues there are 3 biological samples from different pools comprised of 10 animals with a technical replicate for each. Keywords = Mouse sex-specific transcription, gonad specific gene expression Keywords: other

2004-03-28 | GSE1148 | GEO

RNA-sequencing and function analysis of adult gonad tissue identify potential key transcription factors in Macrobrachium resenbergii with sex determination

Project description:M. rosenbergii is a classical aquatic organism with sex-reversed phenomenon by gene-expressed regulation. Reversal of gonad development in M. rosenbergii received great attention but the role of transcription factors for gonadal maintenance and sex determination in adult shrimps remain unclearly. In this study, we sequencing the transcriptomes for the adult testis, ovary and androgenic gland by third-generation sequencing plus second-generation sequencing, and 21,345 were identified. In total, 9,199 differentially expressed genes (DEGs) were identified in pairwise comparison where 2,990 DEGs were upregulated expression in testis than other tissues, and 5,775 DEGs were upregulated expression in ovary, and 2,244 DEGs were upregulated expression in androgenic. There are 272 differentially expressed transcription factors (DETs) in total. The upregulated DETs in one of the testis, ovary and androgenic glands compared to the others were 107, 152 and 13 respectively. The GO and KEGG of DETs analysis suggested that MAPK signaling pathway and transcriptional misregulation pathway have key function in testis, the thyroid hormone signaling pathway and neuroactive ligand-receptor interaction play a important role in ovary. We determined the existence of networks containing significant DETs related to sex determination in the adult goands of M. rosenbergii. The key DETs are Piwi (only expression in testis and ovary) and Argonaute 2 (only expression in ovary) might be involved with the regulation of ovary development.

2021-04-11 | GSE171823 | GEO

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