Lee Tong Y, Yoon Kyoung-Hye, Lee Jin I
Division of Biological Science and Technology, Yonsei University.
Division of Biological Science and Technology, Yonsei University;
J Vis Exp. 2016 Dec 12(118):55048. doi: 10.3791/55048.
The use of genetic model organisms such as Caenorhabditis elegans has led to seminal discoveries in biology over the last five decades. Most of what we know about C. elegans is limited to laboratory cultivation of the nematodes that may not necessarily reflect the environments they normally inhabit in nature. Cultivation of C. elegans in a 3D habitat that is more similar to the 3D matrix that worms encounter in rotten fruits and vegetative compost in nature could reveal novel phenotypes and behaviors not observed in 2D. In addition, experiments in 3D can address how phenotypes we observe in 2D are relevant for the worm in nature. Here, a new method in which C. elegans grows and reproduces normally in three dimensions is presented. Cultivation of C. elegans in Nematode Growth Tube-3D (NGT-3D) can allow us to measure the reproductive fitness of C. elegans strains or different conditions in a 3D environment. We also present a novel method, termed Nematode Growth Bottle-3D (NGB-3D), to cultivate C. elegans in 3D for microscopic analysis. These methods allow scientists to study C. elegans biology in conditions that are more reflective of the environments they encounter in nature. These can help us to understand the overlying evolutionary relevance of the physiology and behavior of C. elegans we observe in the laboratory.
在过去的五十年里,诸如秀丽隐杆线虫等遗传模式生物的使用在生物学领域带来了开创性的发现。我们对秀丽隐杆线虫的了解大多局限于对线虫的实验室培养,而这种培养不一定能反映它们在自然环境中的栖息状况。在更类似于线虫在自然界腐烂果实和植物堆肥中所遇到的三维基质的三维环境中培养秀丽隐杆线虫,可能会揭示出二维培养中未观察到的新表型和行为。此外,三维实验可以解决我们在二维中观察到的表型如何与自然界中的线虫相关的问题。在此,我们提出一种新方法,即秀丽隐杆线虫能在三维环境中正常生长和繁殖。在三维线虫生长管(NGT-3D)中培养秀丽隐杆线虫,能让我们在三维环境中测量秀丽隐杆线虫菌株或不同条件下的繁殖适应性。我们还提出了一种新方法,称为三维线虫生长瓶(NGB-3D),用于在三维环境中培养秀丽隐杆线虫以进行显微镜分析。这些方法使科学家能够在更能反映线虫在自然环境状况的条件下研究秀丽隐杆线虫生物学。这有助于我们理解在实验室中观察到的秀丽隐杆线虫生理和行为的潜在进化相关性。