Genetics of Male Fertility Group, Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain.
Sequentia Biotech SL, Barcelona, Spain.
Mol Hum Reprod. 2021 May 29;27(6). doi: 10.1093/molehr/gaab035.
Transcriptome analyses using high-throughput methodologies allow a deeper understanding of biological functions in different cell types/tissues. The present study provides an RNA-seq profiling of human sperm mRNAs and lncRNAs (messenger and long non-coding RNAs) in a well-characterized population of fertile individuals. Sperm RNA was extracted from twelve ejaculate samples under strict quality controls. Poly(A)-transcripts were sequenced and aligned to the human genome. mRNAs and lncRNAs were classified according to their mean expression values (FPKM: Fragments Per Kilobase of transcript per Million mapped reads) and integrity. Gene Ontology analysis of the Expressed and Highly Expressed mRNAs showed an involvement in diverse reproduction processes, while the Ubiquitously Expressed and Highly Stable mRNAs were mainly involved in spermatogenesis. Transcription factor enrichment analyses revealed that the Highly Expressed and Ubiquitously Expressed sperm mRNAs were primarily regulated by zinc-fingers and spermatogenesis-related proteins. Regarding the Expressed lncRNAs, only one-third of their potential targets corresponded to Expressed mRNAs and were enriched in cell-cycle regulation processes. The remaining two-thirds were absent in sperm and were enriched in embryogenesis-related processes. A significant amount of post-testicular sperm mRNAs and lncRNAs was also detected. Even though our study is solely directed to the poly-A fraction of sperm transcripts, results indicate that both sperm mRNAs and lncRNAs constitute a footprint of previous spermatogenesis events and are configured to affect the first stages of embryo development.
利用高通量方法进行转录组分析可以更深入地了解不同细胞类型/组织中的生物学功能。本研究对来自特征明确的生育个体的人精子 mRNA 和 lncRNA(信使和长非编码 RNA)进行了 RNA-seq 分析。从 12 个精液样本中严格控制质量提取精子 RNA。对 Poly(A)- 转录本进行测序,并与人类基因组进行比对。根据其平均表达值(FPKM:每百万映射读数的转录物每千碱基片段)和完整性对 mRNA 和 lncRNA 进行分类。Expressed 和 Highly Expressed mRNA 的基因本体论分析表明,它们参与了多种生殖过程,而 Ubiquitously Expressed 和 Highly Stable mRNA 主要参与了精子发生。转录因子富集分析表明,Highly Expressed 和 Ubiquitously Expressed 精子 mRNA 主要受锌指和与精子发生相关的蛋白调控。至于 Expressed lncRNA,只有三分之一的潜在靶标对应于 Expressed mRNA,并富集在细胞周期调控过程中。其余三分之二在精子中不存在,富集在胚胎发生相关过程中。还检测到大量的睾丸后精子 mRNA 和 lncRNA。尽管我们的研究仅针对精子转录物的 Poly(A) 部分,但结果表明,精子 mRNA 和 lncRNA 都构成了先前精子发生事件的足迹,并被配置为影响胚胎发育的最初阶段。