Grant W N, Stasiuk S, Newton-Howes J, Ralston M, Bisset S A, Heath D D, Shoemaker C B
AgResearch Ltd, Wallaceville Animal Research Centre, Ward Street, P.O. Box 40063, Upper Hutt, New Zealand.
Int J Parasitol. 2006 Apr;36(4):453-66. doi: 10.1016/j.ijpara.2005.11.009. Epub 2006 Jan 19.
Commonly studied nematode parasites have not proven amenable to simple genetic analyses and this has significantly reduced the available research options. We introduce here a nematode parasite of mammals, Parastrongyloides trichosuri, which has features uniquely suited for genetic analysis. This parasite has the capacity to undergo multiple reproductive cycles as a free-living worm and thereby amplify the numbers of its infective L3s in faeces. Culture conditions are presented that permit facile laboratory maintenance of this worm for >90 free-living life cycles (to date) without the need for re-entry into a permissive host. Even after long maintenance as a free-living worm, culture conditions can be manipulated to favour development of infective L3 worms, which remain able to successfully infect their marsupial hosts. The switch to infective L3 development is triggered by a secreted factor contained in culture medium conditioned by multiple generations of free-living worm culture. It is simple to perform single pair crosses with P. trichosuri to carry out Mendelian genetics in the laboratory and this has been done multiple times with sibling pairs to generate highly inbred lines. Lines of worms can readily be cryopreserved and recovered. Over 7000 expressed sequence tags have been produced from cDNAs at different life cycle stages and used to identify single nucleotide polymorphisms and microsatellites as genetic markers. Free-living worms live only a few days on average while the patency of parasitic infections can last for several months. Since we show this is not the result of re-infection, we conclude that parasitic worms have a lifespan capacity at least 20-30 times longer than their free-living counterparts. We discuss how it should be possible to exploit these unique features of P. trichosuri as a model for future studies that explore the genetic basis of longevity and parasitism.
常见的被研究线虫寄生虫尚未被证明适用于简单的遗传分析,这显著减少了可用的研究选择。我们在此介绍一种哺乳动物的线虫寄生虫——毛首类圆线虫(Parastrongyloides trichosuri),它具有特别适合遗传分析的特征。这种寄生虫能够作为自由生活的蠕虫经历多个繁殖周期,从而在粪便中扩增其感染性L3幼虫的数量。本文介绍了培养条件,在这些条件下,这种蠕虫可以在实验室中轻松维持超过90个自由生活周期(截至目前),而无需重新进入适宜的宿主。即使作为自由生活的蠕虫长期维持培养后,培养条件仍可被操控以促进感染性L3蠕虫的发育,这些蠕虫仍能够成功感染其有袋类宿主。向感染性L3发育的转变是由多代自由生活蠕虫培养条件下的培养基中所含的一种分泌因子触发的。用毛首类圆线虫进行单对杂交以在实验室中进行孟德尔遗传学研究很简单,并且已经用同胞对多次进行了这样的操作以产生高度近交系。线虫品系可以很容易地冷冻保存和复苏。已经从不同生命周期阶段的cDNA中产生了超过7000个表达序列标签,并用于鉴定单核苷酸多态性和微卫星作为遗传标记。自由生活的蠕虫平均仅存活几天,而寄生虫感染的排虫期可持续数月。由于我们表明这不是再次感染的结果,我们得出结论,寄生蠕虫的寿命至少比其自由生活的同类长20 - 30倍。我们讨论了如何利用毛首类圆线虫的这些独特特征作为未来研究的模型,以探索长寿和寄生的遗传基础。