目的探讨人精子DNA甲基化水平与精液常规参数和精子染色质结构完整性的关系。方法纳入2014年重庆市大学生男性生殖健康(male reproductive health in Chongqing college students,MARHCS)队列的73名志愿者,分别采用计算机辅助精子分析系...目的探讨人精子DNA甲基化水平与精液常规参数和精子染色质结构完整性的关系。方法纳入2014年重庆市大学生男性生殖健康(male reproductive health in Chongqing college students,MARHCS)队列的73名志愿者,分别采用计算机辅助精子分析系统(computer-aided sperm analysis,CASA)和精子染色质结构分析(sperm chromatin structure assay,SCSA)系统检测分析精液常规参数和精子染色质结构完整性指标。通过酶联免疫吸附测定法(ELISA)检测精子DNA 5mC含量,反映精子DNA甲基化水平。同时采用调查问卷收集志愿者的年龄、BMI、禁欲时间及生活方式等数据。多元线性回归模型分析精子DNA 5mC含量与精液参数和精子染色质结构完整性的相关性,同时比较高水平和低水平精子DNA 5mC组(按照5mC%中位数二分类)志愿者各级精子百分率的差异。结果志愿者各项精液常规参数(中位数):精液体积为4.18 mL,精子密度为55.60×106/mL,精子总数为246.93×106,精子活力为57.00%;染色质结构完整性:精子DNA碎片化指数(DNA fragmentation index,DFI)和高DNA着色性(high DNA stainability,HDS)指标的中位数分别是10.99%和3.39%;精子DNA 5mC水平的中位数为2.63%。校正混杂因素后,多元线性回归结果显示精子DNA 5mC与精子密度(β=-0.22,95%CI:-0.36~-0.06,P=0.008)和精子总数(β=-0.20,95%CI:-0.33~-0.05,P=0.015)负相关,与精子DFI和HDS指标的相关性分析无统计学意义(P>0.05)。与低水平精子DNA 5mC组相比,高水平精子DNA 5mC组各级精子百分率差异无统计学意义(P>0.05)。结论精子DNA甲基化水平升高可能是精子密度和精子总数下降的标志,而与精子染色质结构完整性指标无关。展开更多
Background:N6-methyladenosine(m^(6)A)and DNA 5-methylcytosine(5mC)methylation plays crucial roles in diverse biological processes,including skeletal muscle development and growth.Recent studies unveiled a potential li...Background:N6-methyladenosine(m^(6)A)and DNA 5-methylcytosine(5mC)methylation plays crucial roles in diverse biological processes,including skeletal muscle development and growth.Recent studies unveiled a potential link between these two systems,implicating the potential mechanism of coordinated transcriptional and post-transcrip-tional regulation in porcine prenatal myogenesis and postnatal skeletal muscle growth.Methods:Immunofluorescence and co-IP assays were carried out between the 5mC writers and m^(6)A writers to investigate the molecular basis underneath.Large-scale in-house transcriptomic data were compiled for applying weighted correlation network analysis(WGCNA)to identify the co-expression patterns of m^(6)A and 5mC regulators and their potential role in pig myogenesis.Whole-genome bisulfite sequencing(WGBS)and methylated RNA immu-noprecipitation sequencing(MeRIP-seq)were performed on the skeletal muscle samples from Landrace pigs at four postnatal growth stages(days 30,60,120 and 180).Results:Significantly correlated expression between 5mC writers and m^(6)A writers and co-occurrence of 5mC and m^(6)A modification were revealed from public datasets of C2C12 myoblasts.The protein-protein interactions between the DNA methylase and the m^(6)A methylase were observed in mouse myoblast cells.Further,by analyzing tran-scriptome data comprising 81 pig skeletal muscle samples across 27 developmental stages,we identified a 5mC/m^(6)A epigenetic module eigengene and decoded its potential functions in pre-or post-transcriptional regulation in postnatal skeletal muscle development and growth of pigs.Following integrative multi-omics analyses on the WGBS methylome data and MeRIP-seq data for both m^(6)A and gene expression profiles revealed a genome/transcriptome-wide correlated dynamics and co-occurrence of 5mC and m^(6)A modifications as a consequence of 5mC/m^(6)A crosstalk in the postnatal myogenesis progress of pigs.Last,we identified a group of myogenesis-related genes collaboratively regulated by both 5mC and m^(6)A modifications in postnatal skeletal muscle growth in pigs.Conclusions:Our study discloses a potential epigenetic mechanism in skeletal muscle development and provides a novel direction for animal breeding and drug development of related human muscle-related diseases.展开更多
基金Project supported by the 111 Project(No.B13026)the National High-Tech R&D Program(863)of China(No.2012AA02A601)+1 种基金the Fundamental Research Funds for the Central Universitiesthe Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents
基金supported by the Agricultural Science and Technology Innovation Program and The Elite Young Scientists Program of CAAS.ZT was supported by the National Natural Science Foundation of China(31830090)the Basic and Applied Basic Research Foundation of Guangdong province(2019B1515120059)+1 种基金the Shenzhen Dapeng New District Special Fund for Industry Development(KY20180114)the Agricultural Science and Technology Innovation Program(CAAS-ZDRW202006).
文摘Background:N6-methyladenosine(m^(6)A)and DNA 5-methylcytosine(5mC)methylation plays crucial roles in diverse biological processes,including skeletal muscle development and growth.Recent studies unveiled a potential link between these two systems,implicating the potential mechanism of coordinated transcriptional and post-transcrip-tional regulation in porcine prenatal myogenesis and postnatal skeletal muscle growth.Methods:Immunofluorescence and co-IP assays were carried out between the 5mC writers and m^(6)A writers to investigate the molecular basis underneath.Large-scale in-house transcriptomic data were compiled for applying weighted correlation network analysis(WGCNA)to identify the co-expression patterns of m^(6)A and 5mC regulators and their potential role in pig myogenesis.Whole-genome bisulfite sequencing(WGBS)and methylated RNA immu-noprecipitation sequencing(MeRIP-seq)were performed on the skeletal muscle samples from Landrace pigs at four postnatal growth stages(days 30,60,120 and 180).Results:Significantly correlated expression between 5mC writers and m^(6)A writers and co-occurrence of 5mC and m^(6)A modification were revealed from public datasets of C2C12 myoblasts.The protein-protein interactions between the DNA methylase and the m^(6)A methylase were observed in mouse myoblast cells.Further,by analyzing tran-scriptome data comprising 81 pig skeletal muscle samples across 27 developmental stages,we identified a 5mC/m^(6)A epigenetic module eigengene and decoded its potential functions in pre-or post-transcriptional regulation in postnatal skeletal muscle development and growth of pigs.Following integrative multi-omics analyses on the WGBS methylome data and MeRIP-seq data for both m^(6)A and gene expression profiles revealed a genome/transcriptome-wide correlated dynamics and co-occurrence of 5mC and m^(6)A modifications as a consequence of 5mC/m^(6)A crosstalk in the postnatal myogenesis progress of pigs.Last,we identified a group of myogenesis-related genes collaboratively regulated by both 5mC and m^(6)A modifications in postnatal skeletal muscle growth in pigs.Conclusions:Our study discloses a potential epigenetic mechanism in skeletal muscle development and provides a novel direction for animal breeding and drug development of related human muscle-related diseases.
