State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu Province, 730046, People's Republic China.
Parasitol Res. 2013 Mar;112(3):1189-95. doi: 10.1007/s00436-012-3250-x. Epub 2013 Jan 10.
Ascaris nematodes, which cause ascariasis in humans and pigs, are among the most important nematodes from both health and economic perspectives. microRNA (miRNA) is now recognized as key regulator of gene expression at posttranscription level. The public availability of the genome and transcripts of Ascaris suum provides powerful resources for the research of miRNA profiles of the parasite. Therefore, we investigated and compared the miRNA profiles of male and female adult A. suum using Solexa deep sequencing combined with bioinformatic analysis and stem-loop reverse transcription polymerase chain reaction. Deep sequencing of small RNAs yielded 11.71 and 11.72 million raw reads from male and female adults of A. suum, respectively. Analysis showed that the noncoding RNA of the two genders, including tRNA, rRNA, snRNA, and snoRNA, were similar. By mapping to the A. suum genome, we obtained 494 and 505 miRNA candidates from the female and male parasite, respectively, and 87 and 82 of miRNA candidates were consistent with A. suum miRNAs deposited in the miRBase database. Among the miRNA candidates, 154 were shared by the two genders, and 340 and 351 were female and male specific with their target numbers ranged from one to thousands, respectively. Functional prediction revealed a set of elongation factors, heat shock proteins, and growth factors from the targets of gender-specific miRNAs, which were essential for the development of the parasite. Moreover, major sperm protein and nematode sperm cell motility protein were found in targets of the male-specific miRNAs. Ovarian message protein was found in targets of the female-specific miRNAs. Enrichment analysis revealed significant differences among Gene Ontology terms of miRNA targets of the two genders, such as electron carrier and biological adhesion process. The regulating functions of gender-specific miRNAs was therefore not only related to the fundamental functions of cells but also were essential to the germ development of the parasite. The present study provides a framework for further research of Ascaris miRNAs, and consequently leads to the development of potential nucleotide vaccines against Ascaris of human and animal health significance.
蛔虫,可引起人类和猪的蛔虫病,是从健康和经济角度来看最重要的线虫之一。microRNA (miRNA) 现在被认为是转录后水平基因表达的关键调节因子。猪蛔虫基因组和转录本的公开可用性为寄生虫 miRNA 谱的研究提供了强大的资源。因此,我们使用 Solexa 深度测序结合生物信息学分析和茎环反转录聚合酶链反应,研究并比较了雌雄成虫 A. suum 的 miRNA 图谱。从雌雄成虫 A. suum 中分别获得了 1171 万和 1172 万条原始读段的小 RNA 深度测序。分析表明,两种性别非编码 RNA,包括 tRNA、rRNA、snRNA 和 snoRNA,相似。通过映射到 A. suum 基因组,我们从雌性寄生虫中获得了 494 个和 505 个 miRNA 候选物,从雄性寄生虫中获得了 87 个和 82 个 miRNA 候选物,与 miRBase 数据库中储存的 A. suum miRNAs 一致。在 miRNA 候选物中,有 154 个是两性共享的,有 340 个和 351 个是雌性和雄性特有的,其靶标数量从一个到数千个不等。功能预测显示,从性别特异性 miRNA 的靶标中发现了一组伸长因子、热休克蛋白和生长因子,这对寄生虫的发育至关重要。此外,在雄性特异性 miRNA 的靶标中发现了主要精子蛋白和线虫精子细胞运动蛋白,在雌性特异性 miRNA 的靶标中发现了卵母细胞信息蛋白。GO 术语富集分析显示,两种性别 miRNA 靶标之间的 GO 术语存在显著差异,如电子载体和生物粘附过程。因此,性别特异性 miRNA 的调节功能不仅与细胞的基本功能有关,而且对寄生虫的生殖发育也至关重要。本研究为进一步研究猪蛔虫 miRNA 提供了框架,进而为针对人类和动物健康重要性的蛔虫的潜在核苷酸疫苗的开发提供了思路。