Wang Xingdong, Pei Jie, Guo Shaoke, Cao Mengli, Kang Yandong, Xiong Lin, La Yongfu, Bao Pengjia, Liang Chunnian, Yan Ping, Guo Xian
Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Key Laboratory of Animal Genetics and Breeding on Tibetan Plateau, Ministry of Agriculture and Rural Affairs, Lanzhou, China.
Front Vet Sci. 2022 Aug 9;9:971515. doi: 10.3389/fvets.2022.971515. eCollection 2022.
N-methyladenosine (mA) is the most common form of eukaryotic mRNA modification, and it has been shown to exhibit broad regulatory activity in yeast, plants, and mammals. The specific role of mA methylation as a regulator of spermatogenesis, however, has yet to be established. In this experiment, through a series of preliminary studies and methylated RNA immunoprecipitation sequencing, the mA map of cattle-yak testicular tissue was established as a means of exploring how mA modification affects cattle-yak male infertility. Cattle-yak testis tissues used in this study were found to contain sertoli cells and spermatogonia. Relative to sexually mature yak samples, those isolated from cattle-yak testis exhibited slightly reduced levels of overall methylation, although these levels were significantly higher than those in samples from pre-sexually mature yaks. Annotation analyses revealed that differentially methylated peaks were most concentrated in exonic regions, with progressively lower levels of concentration in the 3'-untranslated region (UTR) and 5'-UTR regions. To further explore the role of such mA modification, enrichment analyses were performed on differentially methylated and differentially expressed genes in these samples. For the cattle-yaks vs. 18-months-old yaks group comparisons, differentially methylated genes were found to be associated with spermatogenesis-related GO terms related to the cytoskeleton and actin-binding, as well as with KEGG terms related to the regulation of the actin cytoskeleton and the MAPK signaling pathway. Similarly, enrichment analyses performed for the cattle-yaks vs. 5-years-old yaks comparison revealed differentially methylated genes to be associated with GO terms related to protein ubiquitination, ubiquitin ligase complexes, ubiquitin-dependent protein catabolism, and endocytotic activity, as well as with KEGG terms related to apoptosis and the Fanconi anemia pathway. Overall, enrichment analyses for the cattle-yaks vs. 18-months-old yaks comparison were primarily associated with spermatogenesis, whereas those for the cattle-yaks vs. 5-years-old yaks comparison were primarily associated with apoptosis.
N6-甲基腺苷（mA）是真核生物mRNA修饰中最常见的形式，并且已证明其在酵母、植物和哺乳动物中具有广泛的调节活性。然而，mA甲基化作为精子发生调节因子的具体作用尚未确定。在本实验中，通过一系列初步研究和甲基化RNA免疫沉淀测序，建立了牦牛睾丸组织的mA图谱，以此探索mA修饰如何影响牦牛雄性不育。本研究中使用的牦牛睾丸组织含有支持细胞和精原细胞。相对于性成熟牦牛样本，从牦牛睾丸分离的样本整体甲基化水平略有降低，尽管这些水平显著高于性成熟前牦牛样本中的水平。注释分析显示，差异甲基化峰最集中在外显子区域，在3'-非翻译区（UTR）和5'-UTR区域的集中水平逐渐降低。为了进一步探索这种mA修饰的作用，对这些样本中差异甲基化和差异表达的基因进行了富集分析。对于牦牛与18月龄牦牛组的比较，发现差异甲基化基因与细胞骨架和肌动蛋白结合相关的精子发生相关GO术语有关，以及与肌动蛋白细胞骨架调节和MAPK信号通路相关的KEGG术语有关。同样，对牦牛与5岁牦牛比较进行的富集分析显示，差异甲基化基因与蛋白质泛素化、泛素连接酶复合物、泛素依赖性蛋白质分解代谢和内吞活性相关的GO术语有关，以及与细胞凋亡和范可尼贫血途径相关的KEGG术语有关。总体而言，牦牛与18月龄牦牛比较的富集分析主要与精子发生相关，而牦牛与5岁牦牛比较的富集分析主要与细胞凋亡相关。
