Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun, 130062, China.
Laboratory of Veterinary Microbiology, Department of Veterinary Medicine, Yanbian University, Yanji, 133002, China.
Parasitol Res. 2021 Jun;120(6):2165-2174. doi: 10.1007/s00436-021-07155-2. Epub 2021 Apr 24.
Neospora caninum is an important pathogen commonly causing spontaneous abortion in livestock. The parasite is known to remain in cysts in an inactive state; or it can undergo expansive development within an intermediate host. However, the mechanisms that trigger the proliferation of N. caninum have not been thoroughly elucidated. For various organisms, it has been demonstrated that microRNAs (miRNAs) can act as important endogenous regulatory factors in gene regulation during cell differentiation and development. However, miRNAs and their function have not been studied in N. caninum. In this study, small RNA libraries from N. caninum tachyzoites (NC-1 strain) were analyzed using a high-throughput RNA sequencing technology combined with systematic bioinformatics analysis. A considerable number of novel miRNAs from N. caninum NC-1 strain tachyzoites were identified. Of the 300 miRNAs found by bioinformatics analysis, 10 were conserved miRNAs belonging to 10 metazoan miRNA families, while 290 were novel miRNAs. The expression of 13 novel miRNAs was verified by real-time quantitative PCR (qRT-PCR). Data from this study provided and identified authentic miRNAs for the first time in N. caninum. The study also introduces a framework for further investigations of RNAi-dependent regulatory mechanisms of the parasite and provides data for further understanding of N. caninum development.
刚地弓形虫是一种重要的病原体，常导致家畜自然流产。该寄生虫已知以囊包的形式处于非活跃状态；或者在中间宿主中进行广泛的发育。然而，触发刚地弓形虫增殖的机制尚未被彻底阐明。对于各种生物体，已经证明 microRNAs（miRNAs）可以在细胞分化和发育过程中的基因调控中作为重要的内源性调节因子发挥作用。然而，miRNAs 及其功能在刚地弓形虫中尚未得到研究。在这项研究中，使用高通量 RNA 测序技术结合系统生物信息学分析，对刚地弓形虫速殖子（NC-1 株）的小 RNA 文库进行了分析。从 NC-1 株速殖子中鉴定出大量新的刚地弓形虫 miRNA。通过生物信息学分析发现的 300 个 miRNA 中，有 10 个是属于 10 个后生动物 miRNA 家族的保守 miRNA，而 290 个是新的 miRNA。通过实时定量 PCR（qRT-PCR）验证了 13 个新 miRNA 的表达。本研究首次为刚地弓形虫提供并鉴定了真实的 miRNA。该研究还为进一步研究寄生虫的 RNAi 依赖调节机制提供了框架，并为进一步了解刚地弓形虫的发育提供了数据。
