Deline Marshall L, Vrablik Tracy L, Watts Jennifer L
School of Molecular Biosciences, Washington State University.
J Vis Exp. 2013 Nov 29(81):50879. doi: 10.3791/50879.
Fatty acids are essential for numerous cellular functions. They serve as efficient energy storage molecules, make up the hydrophobic core of membranes, and participate in various signaling pathways. Caenorhabditis elegans synthesizes all of the enzymes necessary to produce a range of omega-6 and omega-3 fatty acids. This, combined with the simple anatomy and range of available genetic tools, make it an attractive model to study fatty acid function. In order to investigate the genetic pathways that mediate the physiological effects of dietary fatty acids, we have developed a method to supplement the C. elegans diet with unsaturated fatty acids. Supplementation is an effective means to alter the fatty acid composition of worms and can also be used to rescue defects in fatty acid-deficient mutants. Our method uses nematode growth medium agar (NGM) supplemented with fatty acid sodium salts. The fatty acids in the supplemented plates become incorporated into the membranes of the bacterial food source, which is then taken up by the C. elegans that feed on the supplemented bacteria. We also describe a gas chromatography protocol to monitor the changes in fatty acid composition that occur in supplemented worms. This is an efficient way to supplement the diets of both large and small populations of C. elegans, allowing for a range of applications for this method.
脂肪酸对众多细胞功能至关重要。它们是高效的能量储存分子，构成细胞膜的疏水核心，并参与各种信号通路。秀丽隐杆线虫能合成一系列ω-6和ω-3脂肪酸所需的所有酶。这一点，再加上其简单的解剖结构和可用的多种遗传工具，使其成为研究脂肪酸功能的一个有吸引力的模型。为了研究介导膳食脂肪酸生理效应的遗传途径，我们开发了一种用不饱和脂肪酸补充秀丽隐杆线虫食物的方法。补充是改变线虫脂肪酸组成的有效手段，也可用于挽救脂肪酸缺乏突变体的缺陷。我们的方法使用补充了脂肪酸钠盐的线虫生长培养基琼脂（NGM）。补充平板中的脂肪酸会掺入细菌食物源的膜中，然后被以补充细菌为食的秀丽隐杆线虫摄取。我们还描述了一种气相色谱法方案，以监测补充线虫中发生的脂肪酸组成变化。这是一种补充大小群体秀丽隐杆线虫食物的有效方法，使该方法有一系列应用。
