Margie Olivia, Palmer Chris, Chin-Sang Ian
Life Sciences, Queen's University.
J Vis Exp. 2013 Apr 27(74):e50069. doi: 10.3791/50069.
Many organisms use chemotaxis to seek out food sources, avoid noxious substances, and find mates. Caenorhabditis elegans has impressive chemotaxis behavior. The premise behind testing the response of the worms to an odorant is to place them in an area and observe the movement evoked in response to an odorant. Even with the many available assays, optimizing worm starting location relative to both the control and test areas, while minimizing the interaction of worms with each other, while maintaining a significant sample size remains a work in progress (1-10). The method described here aims to address these issues by modifying the assay developed by Bargmann et al.(1). A Petri dish is divided into four quadrants, two opposite quadrants marked "Test" and two are designated "Control". Anesthetic is placed in all test and control sites. The worms are placed in the center of the plate with a circle marked around the origin to ensure that non-motile worms will be ignored. Utilizing a four-quadrant system rather than one 2 or two 1 eliminates bias in the movement of the worms, as they are equidistant from test and control samples, regardless of which side of the origin they began. This circumvents the problem of worms being forced to travel through a cluster of other worms to respond to an odorant, which can delay worms or force them to take a more circuitous route, yielding an incorrect interpretation of their intended path. This method also shows practical advantages by having a larger sample size and allowing the researcher to run the assay unattended and score the worms once the allotted time has expired.
许多生物体利用化学趋向性来寻找食物来源、避开有害物质并寻找配偶。秀丽隐杆线虫具有令人印象深刻的化学趋向性行为。测试线虫对气味剂反应的前提是将它们放置在一个区域,并观察它们对气味剂产生的运动反应。即使有许多可用的检测方法,但要优化线虫相对于对照区域和测试区域的起始位置,同时尽量减少线虫之间的相互作用,并保持足够大的样本量,仍在不断改进中(1 - 10)。这里描述的方法旨在通过改进Bargmann等人开发的检测方法(1)来解决这些问题。将一个培养皿分成四个象限,两个相对的象限标记为“测试”,另外两个指定为“对照”。在所有测试和对照位置都放置麻醉剂。线虫被放置在培养皿的中心,在原点周围画一个圈,以确保不动的线虫被忽略。使用四象限系统而不是一个2或两个1的系统可以消除线虫运动中的偏差,因为无论它们从原点的哪一侧开始,它们与测试样本和对照样本的距离都是相等的。这避免了线虫被迫穿过一群其他线虫去响应气味剂的问题,这可能会使线虫延迟或迫使它们采取更迂回的路线,从而对它们的预期路径产生错误的解读。这种方法还具有实际优势,即样本量更大,并且允许研究人员在无人值守的情况下进行检测,并在规定时间结束后对线虫进行评分。