摘要
目的探讨早老素在日本血吸虫生殖发育及产卵中的作用。方法分别收集日本血吸虫尾蚴感染小鼠后16、18、24 d后虫体,首先通过荧光定量PCR分析早老素在上述3个阶段表达;从感染后28 d雌虫中分离卵巢,荧光定量PCR分析早老素在卵巢的表达量。体外培养28 d雌雄合抱的成虫,应用siRNA对早老素进行干扰,10 d后,观察早老素被敲降后对血吸虫产卵量,雌雄合抱及生殖系统的影响。结果荧光定量PCR结果显示,早老素在24 d雌虫表达量明显上调,同时证实该基因在卵巢组织高表达。当早老素被敲降后,雌雄合抱率和雌虫的产卵量也有明显下降。激光共聚焦结果显示相对于对照组和Mock组,RNA干扰组的生殖细胞减少,生殖系统形态出现异常。结论早老素在日本血吸虫生殖发育和产卵中起重要调控作用。
Objective To study the role of presenilin in the reproductive development and oviposition of Schistosoma japonicum(S.japonicum).Methods The 16,18,24 or 28 days post infection(dpi)worms were collected from the mice infected with S.japonicum,then the expression levels of presenilin in the 16,18 or 24dpi S.japonicum females were analyzed by real-time quantitative reverse transcription PCR(qRT-PCR);The ovaries were collected from 28 dpi S.japonicum females,then the expression levels of presenilin in ovary were analyzed by qRT-PCR.The paired schistosomes of 28 dpi were transfected with specific presenilin siRNA and cultured in vitro for 10 days,then the morphological changes in reproductive organs,male-female pairing and egg production were observed after presenilin gene knockdown.Results The presenilin mRNA expression levels in females 24 dpi were markedly higher than those in females 18 dpi or 16 dpi,in both male and female parasites,higher amounts of the presenilin gene were expressed in ovary tissues.The RNAi of 28 days'pairing worms were treated with siRNA,after 10 days of culture in vitro,the results showed that compared with the control group and the Mock group,the male and female pairing rate of the experimental group decreased significantly;the results of confocal laser scanning microscope showed that in the interference group,the morphology of the reproductive system changed,the number of germ cells decreased and the morphology was abnormal.Conclusion Presenilin plays an important role in the reproductive development and oviposition of S.japonicum.
作者
杨旭晗
杨菲
沈际佳
刘淼
Yang Xuhan;Yang Fei;Shen Jijia(Dept of Microbiology and Parasitology,Anhui Provincial Laboratory of Pathogen Biology and Anhui Key Laboratory of Zoonoses,School of Basic Medicine,Anhui Medical University,Hefei 230032)
出处
《安徽医科大学学报》
CAS
北大核心
2021年第10期1590-1594,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81271865)
安徽高校自然科学研究项目(编号:KJ2016A333)。
关键词
日本血吸虫
早老素
RNA干扰
生殖
Schistosoma japonicum
presenilin
RNA interference
reproduction
