Label-free quantitative analysis of lipid metabolism in living Caenorhabditis elegans
Label-free quantitative analysis of lipid metabolism in living Caenorhabditis elegans
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DOI：
10.1194/jlr.d000638
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发表时间：
2010-03-01
影响因子：
6.5
通讯作者：
Cheng, Ji-Xin
中科院分区：
文献类型：
--
作者：
Le, Thuc T.;Duren, Holli M.;Cheng, Ji-Xin
脂质在生物结构和功能中的普遍存在表明脂质代谢受到高度调节。然而，目前脂质研究的侵入性技术阻止了各种脂质代谢途径之间的动态相互作用的表征。在这里，我们描述了一种非侵入性的方法来研究脂质代谢，使用多功能相干反斯托克斯拉曼散射（汽车）显微镜。使用活的秀丽隐杆线虫作为模式生物，我们报告共存的中性和自发荧光脂质物种的无标记可视化。我们发现中性和自发荧光脂质的相对表达水平可以用来分析突变体C.在编码脂质合成转录因子、LDL受体、转化生长因子β受体、脂质去饱和酶和抗氧化酶的基因中缺失的线虫。此外，通过汽车与指纹共聚焦拉曼分析相结合，我们分析了野生型和突变型C。优雅的。我们的研究表明，复杂的基因型-表型之间的关系，脂质储存，过氧化和去饱和，可以快速和定量分析在一个单一的生活C。优雅。Le，T. T.，H. M.杜伦，M. N. Slipchenko，C-D。Hu和J-X.程活体秀丽隐杆线虫脂质代谢的无标记定量分析。J. Lipid Res. 2010. 51：672-677。
The ubiquity of lipids in biological structures and functions suggests that lipid metabolisms are highly regulated. However, current invasive techniques for lipid studies prevent characterization of the dynamic interactions between various lipid metabolism pathways. Here, we describe a noninvasive approach to study lipid metabolisms using a multifunctional coherent anti-Stokes Raman scattering (CARS) microscope. Using living Caenorhabditis elegans as a model organism, we report label-free visualization of coexisting neutral and autofluorescent lipid species. We find that the relative expression level of neutral and autofluorescent lipid species can be used to assay the genotype-phenotype relationship of mutant C. elegans with deletions in the genes encoding lipid synthesis transcription factors, LDL receptors, transforming growth factor beta receptors, lipid desaturation enzymes, and antioxidant enzymes. Furthermore, by coupling CARS with fingerprint confocal Raman analysis, we analyze the unsaturation level of lipids in wild-type and mutant C. elegans. Our study shows that complex genotype-phenotype relationships between lipid storage, peroxidation, and desaturation can be rapidly and quantitatively analyzed in a single living C. elegans.-Le, T. T., H. M. Duren, M. N. Slipchenko, C-D. Hu, and J-X. Cheng. Label-free quantitative analysis of lipid metabolism in living Caenorhabditis elegans. J. Lipid Res. 2010. 51: 672-677.