Department of Microbiology and Parasitology, School of Basic Medical Sciences, Anhui Medical University, 81# Meishan Road, Hefei, 230032, Anhui, People's Republic of China.
Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, 81# Meishan Road, Hefei, 230032, Anhui, People's Republic of China.
Parasitol Res. 2020 Feb;119(2):501-511. doi: 10.1007/s00436-019-06467-8. Epub 2020 Jan 2.
The elimination of schistosomiasis, a parasitic disease caused by Schistosoma and a major source of morbidity and mortality in developing countries, faces serious challenges. Although the pumilio protein regulates the reproductive organ development in many species, its role in Schistosoma japonicum is unknown. Thus, this study investigated the function of pumilio in S. japonicum reproduction. The complete coding sequences of S. japonicum Pumilio1 (SjPum1) and SjPum2 genes were cloned and characterized. The full-length open-reading frame SjPum1 (2613 nucleotides) and SjPum2 (4479 nucleotides) genes were obtained. Bioinformatics analysis showed that those genes belonged to the PUF (pumilio and FBF) family. Quantitative polymerase chain reaction analyses revealed that SjPum1 and SjPum2 were differentially expressed throughout the S. japonicum life cycle and were highly expressed in reproductive organs. In situ hybridization results showed that mRNA expression of SjPum2 was higher than that of SjPum1 in the ovary and testis. Knocking down SjPum2 using RNA interference techniques to explore potential reproductive functions showed that compared with the control (untransfected or scrambled mRNA-transfected) worms, the morphology of both male and female reproductive organs was altered, the number of eggs produced by paired females was significantly decreased, and the transcription levels of caspase 3 and caspase 7 genes related to apoptosis were significantly increased. The transcription level of Nanos1 gene which related to reproduction was also significantly increased. Therefore, SjPum2 may play a role in the reproductive development of S. japonicum.
血吸虫病是一种由血吸虫引起的寄生虫病,是发展中国家发病率和死亡率的主要原因,消除血吸虫病面临着严峻的挑战。尽管 pumilio 蛋白在许多物种中调节生殖器官的发育,但它在日本血吸虫中的作用尚不清楚。因此,本研究探讨了 pumilio 在日本血吸虫生殖中的作用。克隆和鉴定了日本血吸虫 pumilio1(SjPum1)和 SjPum2 基因的完整编码序列。获得了全长开放阅读框 SjPum1(2613 个核苷酸)和 SjPum2(4479 个核苷酸)基因。生物信息学分析表明,这些基因属于 PUF(pumilio 和 FBF)家族。定量聚合酶链反应分析显示,SjPum1 和 SjPum2 在日本血吸虫生命周期中差异表达,在生殖器官中高度表达。原位杂交结果显示,SjPum2 在卵巢和睾丸中的 mRNA 表达均高于 SjPum1。利用 RNA 干扰技术敲低 SjPum2 以探索潜在的生殖功能,结果显示与对照组(未转染或转染乱序 mRNA)相比,雌雄生殖器官的形态发生改变,配对雌虫产生的卵数显著减少,与凋亡相关的 caspase 3 和 caspase 7 基因的转录水平显著增加。与生殖有关的 Nanos1 基因的转录水平也显著增加。因此,SjPum2 可能在日本血吸虫的生殖发育中发挥作用。