Expression of hepatitis B virus surface antigens induces defective gonad phenotypes in Caenorhabditis elegans

• 作者: Li-Wei Lee Szecheng J Lo Wei-Ning Hong Yi-Yin Chen
• 来源期刊: 世界病毒学杂志
• 学科: 医学
• 关键词: Hepatitis B virus CAENORHABDITIS elegans Green fluorescence proteins Endoplasmic reticulum stress GONAD RETARDATION Surface ANTIGENS
• 年，卷（期）: 2017,（1）
• 页码范围: 17-25
• 页数: 9页
• 分类号: R

摘要

AIM To test whether a simple animal,Caenorhabditis elegans(C.elegans),can be used as an alternative model to study the interaction between hepatitis B virus antigens(HBsAg)and host factors.METHODS Three plasmids that were able to express the large,middle and small forms of HBsAgs(LHBsAg,MHBsAg,and SHBsAg,respectively)driven by a ubiquitous promoter(fib-1)and three that were able to express SHBsAg driven by different tissue-specific promoters were constructed and microinjected into worms.The brood size,egglaying rate,and gonad development of transgenic worms were analyzed using microscopy.Levels of mRNA related to endoplasmic reticulum stress,enpl-1,hsp-4,pdi-3 and xbp-1,were determined using reverse transcription polymerase reaction(RT-PCRs)in three lines of transgenic worms and dithiothreitol(DTT)-treated wild-type worms.RESULTS Severe defects in egg-laying,decreases in brood size,and gonad retardation were observed in transgenic worms expressing SHBsAg whereas moderate defects were observed in transgenic worms expressing LHBsAg and MHBsAg.RT-PCR analysis revealed that enpl-1,hsp-4 and pdi-3 transcripts were significantly elevated in worms expressing LHBsAg and MHBsAg and in wildtype worms pretreated with DTT.By contrast,only pdi-3 was increased in worms expressing SHBsAg.To further determine which tissue expressing SHBsAg could induce gonad retardation,we substituted the fib-1 promoter with three tissue-specific promoters(myo-2 for the pharynx,est-1 for the intestines and mec-7 for the neurons)and generated corresponding transgenic animals.Moderate defective phenotypes were observed in worms expressing SHBsAg in the pharynx and intestines but not in worms expressing SHBsAg in the neurons,suggesting that the secreted SHBsAg may trigger a cross-talk signal between the digestive track and the gonad resulting in defective phenotypes.CONCLUSION Ectopic expression of three forms of HBsAg that causes recognizable phenotypes in transgenic worms suggests that C.elegans can be used as an alternative model for studying vi