Epigenome analysis of Chinese Han male semen samples
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ABSTRACT: Genome wide DNA methylation profiling of Chinese Han male semen samples. The Illumina Human Methylation 450 Beadchip was used to obtain DNA methylation profiles across approximately 450,000 CpGs. Samples included 3 young males (aged 24, 25 and 25 years old) and 3 aged males (aged 63, 60 and 55 years old).
Project description:In this study, 56 semen samples were collected from healthy Korean males aged 18 to 70 and data was obtained utilizing the Illumina Infinium MethylationEPIC BeadChip array (Illumina, San Diego, CA, USA) at Macrogen Inc. (Macrogen, Seoul, Korea).
Project description:<h4><strong>BACKGROUND: </strong>Semen quality is negatively correlated with male age and is mainly quantified by a routine semen analysis, which is descriptive and inconclusive. Sperm proteins or semen metabolites are used as the intermediate or end-products, reflecting changes in semen quality, and hold much promise as a new biomarker to predict fertility in advanced-aged males.</h4><h4><strong>OBJECTIVES: </strong>In this study, we sought to assess whether the semen metabolome and proteome of aged males can affect semen quality and serve as biomarkers for predicting semen quality.</h4><p><strong>MATERIALS AND METHODS: </strong>We retrospectively analyzed 12825 males that underwent semen routine analysis to understand the age-dependent changes in sperm quality. To identify the difference between aged and young adults, metabolomics (n=60) analyses of semen and proteomics (n=12) analyses of sperm were conducted. Finally, integrated machine learning of metabolomics was conducted to screen biomarkers to identify aging semen.</p><p><strong>RESULTS:</strong> We discovered that male age was positively correlated with sperm concentration as well as DNA fragmentation index(DFI), and negatively with progressive motile sperm count, total sperm count, sperm volume and progressive sperm motility. The differential metabolites were significantly enriched in various metabolic pathways, and four of these differential metabolites (Pipamperone, 2,2-Bis(hydroxymethyl)-2,2',2''-nitrilotriethanol, Arg-Pro and Triethyl phosphate) were utilized to establish a biomarker panel to identify aging semen. Proteomic analysis showed that differential proteins were significantly enriched in protein digestion and absorption and some energy-related pathways. An integrated analysis of the metabolome and proteome identified differential energy metabolism and oxidative stress-related proteins, which could explain the decreased motility and the increased DFI of aging sperm.</p><p><strong>DISCUSSION AND CONCLUSION:</strong> We provide compelling evidence that the changes in semen metabolome and sperm proteome are related to the decline of semen quality in aged males. Moreover, a biomarker panel based on four metabolites was established to identify aging semen.</p>
Project description:DNA methylation analysis using the Infinium MethylationEPIC BeadChip arrays was applied to 40 semen-derived DNA samples (age range 24 – 58 years). The aim of the study was to discover differentially methylated sites (DMS) that correlate with age.
Project description:Age-related CpG (AR-CpG) sites are currently the most promising molecular markers for age estimation. However, AR-CpG sites for the Han Chinese population have not yet been systematically identified. Besides, there may be high-performance AR-CpG sites beyond the coverage of the commonly used Illumina HumanMethylation450 BeadChip that covers over 450,000 CpG sites (450K), which accounts only 1.6% of CpG sites in the human genome. Therefore, we initiated a project to perform genome-wide methylation analysis on whole blood samples from 42 healthy Han Chinese individuals (21 males and 21 females, 18–62 years) using the newly developed Illumina Infinium MethylationEPIC array that assesses over 850,000 CpG sites (850K) to identify AR-CpG sites for forensic use systematically. The 850K methylation array contains > 90% of 450K CpG sites but adds 413,745 CpG sites. As expected, we not only replicated several high-performance AR-CpG sites previously discovered using the Illumina 450K BeadChip, such as cg16867656 (ELOVL2), cg22454769 (FHL2), and cg10501210 (C1orf132), but also identified 41 (60.3%), 34 (22.5%), and 382 (48.7%) novel AR-CpG sites from the methylation profiles of 21 males, 21 females, and all individuals, respectively.
Project description:The ability to predict tissue type and donor’s age from molecular profiles of crime scene samples has practical implications in forensics. In order to identify body fluid- and age-associated DNA methylation changes, genome-wide DNA methylation profiling was carried out for body fluids including blood, saliva, semen, menstrual blood, and vaginal fluid obtained from individuals aged 20 to 59. The Illumina Infinium 450k Human DNA methylation Beadchip was used to obtain DNA methylation profiles across approximately 450,000 CpGs in bisulfite converted DNA. Samples included 12 of each blood, saliva and semen samples from 18 male donors aged 20 to 59, and 3 of each vaginal fluid and menstrual blood samples from 4 female donors in their twenties. Genome-wide DNA methylation profiling of body fluids obtained from young and old individuals. The Illumina Infinium 450K Human DNA methylation Beadchip was used to obtain DNA methylation profiles across approximately 450K CpGs from human body fluids including blood, saliva, semen, vaginal fluid and menstrual blood. Bisulfite converted DNA from the 24 samples were hybridised to the Illumina Infinium 450k Human Methylation Beadchip
Project description:The semen of Small Tail Han Sheep has characteristics of high yield, high density, and good motility. To reveal the key miRNAs, mRNAs and miR-Targets regulatory mechanisms in Small Tail Han Sheep testes development, integrated analysis of miRNA and mRNA expression profiles in 2-, 6-, and 12-month-old testes were investigated by RNA-seq technology and bioinformatics methods. As the results shown: 630, 102, and 322 differentially expressed (DE) mRNAs; 5, 1 and 4 DE known miRNAs; 132, 105 and 24 DE novel miRNAs were identified in 2- vs 6-month-old, 6- vs 12-month-old, and 2- vs 12-month-old testes, respectively. GO and pathway analysis showed: in 2- vs 6-month-old testes, DE mRNAs were mainly involved in sexual maturation process and the DE mRNAs were mainly involved in multiple metabolism and biosynthesis pathways; in 6- vs 12-month-old testes, DE mRNAs were mainly involved in metabolism and translation processes, and the most significant pathway that DE mRNAs involved in was ribosome pathway; in 2- vs 12-month-old testes, DE mRNAs were mainly involved in metabolism and physiological processes, and DE mRNAs were mainly involved in multiple metabolism and biosynthesis pathways. Subsequently, 76, 11 and 1 DE miR-Targets were identified in 2- vs 6-month-old, 2- vs 12-month-old, and 6- vs 12-month-old testes, respectively. 3 miR-Target regulatory networks were constructed based on these miR-Targets, which helped to elucidate the regulatory metabolism in Small Tail sheep testes development. Finally, 6 miRNAs and 7 mRNAs were selected to validate the RNA-seq data by RT-PCR.
Project description:In our study, differential male nucleus events and development behaviors were revealed from the fertilized eggs in response to the sperm from males of genotypic sex determination (GSD) and temperature-dependent sex determination (TSD) in gibel carp. When the eggs of maternal fish were fertilized by the sperm from males of GSD, the fertilized egg encountered similar sexual reproduction events and behaviors. However, when the eggs of maternal fish were fertilized by the sperm from males of TSD, a typical process of gynogenesis was observed. To reveal the underlying molecular mechanism of differential sperm nucleus development behaviors in the fertilized eggs, iTRAQ-based quantitative semen proteomics were performed on three semen samples from three males of GSD and three semen samples from three males of TSD respectively.