Characterization of m6A in domesticated yak testis before and after sexual maturity
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ABSTRACT:
PROVIDER: PRJNA751370 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15318976_1.fastq.gz | Fastqsanger.gz | |||
| SRR15318976_2.fastq.gz | Fastqsanger.gz | |||
| SRR15318977_1.fastq.gz | Fastqsanger.gz | |||
| SRR15318977_2.fastq.gz | Fastqsanger.gz | |||
| SRR15318978_1.fastq.gz | Fastqsanger.gz |
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Project description:N6-methyladenosine (m6A) is the most common internal modification of eukaryotic mRNA, and its potential regulatory role has recently been identified in mammals, plants, and yeast. However, little is known about the role of m6A methylation in mammalian sexual maturation. In this study, the testicular tissue of yaks before and after sexual maturation was taken as the research object, and the m6A map of testicular tissue of yaks before and after sexual maturation was obtained through the preliminary experiment and MERIP-seq. The results showed that only spermatogonial cells and some primary spermatocytes could be seen in the testicular tissue of yak before sexual maturation, while spermatogenic cells at different stages could be seen in the testicular tissue after sexual maturation. It was found that the methylation level was higher after sexual maturation than before by detecting the expression of methylation related enzymes and the overall methylation level of testis. There were 1438 differential methylation peaks before and after sexual maturation, of which 1226 were significantly up-regulated and 212 were significantly down-regulated after sexual maturation. Through annotation, it was found that the differential methylation peaks were mainly concentrated in the exon region, followed by 3'UTR, and finally 5'UTR. KEGG pathway analysis showed that Homologous recombination, Notch signaling pathway, Growth hormone synthesis and other signaling pathways may be related to testicular development and maturation. m6A modification was positively correlated with mRNA abundance, suggesting that m6A plays a regulatory role in mammalian sexual maturation. In this study, we reported for the first time the m6A transcriptional map of yak testis, providing a starting point for revealing the function of m6A that may affect the development of yak testis.
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Project description:Testis is the most important male reproductive organ, and the integrity of its physiological function is crucial to the successful production of sperm. In this study, the expression profiles of 11 991 and 8 930 cells in testicular tissue of yak and cattle-yak after sexual maturity were established using Single-cell RNA sequencing. The identification results of cell subpopulations and marker genes were analyzed and their possible mechanisms were predicted.
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Project description:Studying spermatogenesis is the key to understanding the development mechanism of the yak reproductive system. Although N6-methyladenosine (m6A) RNA modification has been reported to regulate spermatogenesis and reproductive function in mammals, the molecular mechanism of m6A in yak testis development and spermatogenesis remains largely unknown. Therefore, we collected testicular tissue from junile and adult yaks, and found that the m6A level significantly increased after sexual maturity in yak. In MeRIP-seq, 1,702 hypermethylated peaks and 724 hypomethylated peaks were identified. The hypermethylated differentially methylated RNAs (DMRs) (CIB2, AK1, FOXJ2, PKDREJ, SLC9A3, and TOPAZ1) mainly regulated spermatogenesis, while the hypomethylated DMRs (BCL6, USP25, CD96, EPHA2, and TAF12) mainly participated in immune response. Functional enrichment analysis showed that DMRs were significantly enriched in the adherens junction, gap junction, and Wnt, PI3K, and mTOR signaling pathways, regulating cell development, spermatogenesis, and testicular endocrine function. The functional analysis of differentially expressed genes showed that they were involved in the biological processes of mitosis, meiosis, and flagellated sperm motility during sexual maturity of yak testis. We also screened the key regulatory factors of testis development and spermatogenesis by combined analysis, which included BRCA1, CREBBP, STAT3, and SMAD4. This study has important research value and practical guiding significance for yak genetic improvement, particularly in understanding the molecular mechanism and evolutionary dynamics of the progression of testis developmental stages in yaks.
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Project description:The potential regulatory role of N6-methyladenosine (m6A), the most prominent mRNA modification in eukaryotes, has recently been identified in mammals, plants, and yeast. However, whether and how m6A methylation is involved in sexual maturation in mammals remains largely unexplored. In this study, testicular tissue was obtained from yaks before and after sexual maturation, and m6A maps were generated via preliminary experiments and methylated RNA immunoprecipitation sequencing. Only spermatogonial cells and a few primary spermatocytes were observed in the testicular tissue of yaks before sexual maturation, while spermatogenic cells at different stages of maturity could observed after sexual maturation. Experiments examining the expression of methylation-related enzymes and overall methylation levels showed that the methylation levels in yak testes increased after sexual maturation. Overall, 1,438 methylation peaks were differentially expressed before and after sexual maturation; 1,226 showed significant up-regulation and 212 showed significant down-regulation after sexual maturation. Annotation analysis showed that the differential methylation peaks were most commonly concentrated in the exon region, followed by the 3'UTR and finally the 5'UTR region. KEGG pathway analysis demonstrated that homologous recombination, the Notch signaling pathway, growth hormone synthesis, and other signaling pathways may be involved in testicular development and maturation in yaks. Levels of most m6A modifications were positively correlated with mRNA abundance, suggesting that m6A plays a regulatory role in mammalian sexual maturation. To our knowledge, this is the first report of an m6A transcriptional map of the yak testes, and our study lays the foundation for elucidating the function of m6A in the development of yak testes.
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Project description:This study used yak and cattle-yak testes from different developmental stages as materials to construct a complete translation map of the testes, and integrated transcriptome and translation results to explore gene expression changes during the sexual maturation process of yak testes. This study utilized Ribo seq technology to construct a transcriptome map of yak testicular development, revealing that the expression of genes related to spermatogenesis is specifically translated and regulated at different developmental stages. In addition, many unknown open reading frames (ORFs) in the testes have been newly identified.
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Project description:Epididymis development is the basis of male reproduction and is a crucial site where sperm maturation occurs. In order to further understand the epididymal development of yak and how to regulate sperm maturation, we conducted a multi-omics analysis. We detected 2274 differential genes, 222 differential proteins and 117 co-expression genes in the cauda epididymis of yak before and after sexual maturity by RNA-seq and proteomics techniques, which included TGFBI, COL1A1, COL1A2, COL3A1, COL12A1, SULT2B1, KRT19, and NPC2. These high abundance genes are mainly related to cell growth, differentiation, adhesion and sperm maturation, and are mainly enriched via extracellular matrix receptor interaction, protein differentiation and absorption, and lysosome and estrogen signaling pathways. The abnormal expression of these genes may lead to the retardation of epididymal cauda development and abnormal sperm function in yak. In conclusion, through single and combined analysis, we provided a theoretical basis for the development of the yak epididymal cauda, sperm maturation, and screening of key genes involved in the regulation of male yak reproduction.
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Project description:RNA sequence analysis identified a total of 8473 differentially expressed genes (DEGs) (3580 up-regulated, 4893 down-regulated), and GO annotation and KEGG enrichment of differential genes found that DEGS were mostly related to spermatogenesis and apoptosis. Among them, Spermatogonia stem cell (SSCs) marker genes (ITGA5, Gfra1, CD9, SOHLH1, SALL4, ID4 and FOXO1) were significantly up-regulated, differentiation spermatogenic cell marker genes (KIT, UCHL1, Ccna1, TNP1 and TXNDC2) were significantly down-regulated, and genes involved in apoptosis (Fas, CTSK, CTSB, CTSC, CTSO, caspase 3, caspase6, and caspase) were significantly up-regulated. Furthermore, the AS events in cattle-yak are significantly lower than in yak, we speculate that lack of mRNA and protein subtypes may lead to spermatogenic arrest in yak testicle.
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Project description:N6-methyladenosine (m6A) is the most prominent mRNA modification in eukaryotes, and its potential regulatory role has recently been identified in mammals, plants, and yeast. However, how m6A methylation regulates spermatogenesis remains unknown. In this study, cattle-yak testis tissue was used as the experimental material, and the m6A map was generated through preliminary experiments and methylated RNA immunoprecipitation sequencing. Only spermatogonia and Sertoli cells were observed in cattle-yak testis tissue. Experiments examining the expression of methylation-related enzymes and the overall methylation level showed that the methylation level in the testis of the cattle-yak was slightly lower than that of the sexually mature yak, but significantly higher than that of the pre-sexually mature yak. Annotation analysis indicated that differentially methylated peaks were most frequently concentrated in exonic regions, followed by 3'UTR and finally 5'UTR regions. Through enrichment analysis of differentially expressed genes and differentially methylated corresponding genes, GO analysis of T-vs-Y group mainly involved spermatogenesis, including cytoskeleton, actin binding, etc. KEGG analysis showed that the differential genes were mainly enriched in actin cytoskeleton regulation and MAPK signaling pathway. GO analysis of the T-vs-M group mainly involved protein ubiquitination, ubiquitin ligase complexes, ubiquitin-dependent protein catabolism and endocytosis. KEGG analysis mainly involved apoptosis and Fanconi anemia pathways. This study will lay the foundation for elucidating the molecular mechanism of m6A in male sterility of cattle-yak.
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