Abstract:
Stable isotope-resolved metabolomics comprises a critical set of technologies that can be applied to a wide variety of systems, from isolated cells to whole organisms, to define metabolic pathway usage and responses to perturbations such as drugs or mutations, as well as providing the basis for flux analysis. As the diversity of stable isotope-enriched compounds is very high, and with newer approaches to multiplexing, the coverage of metabolism is now very extensive. However, as the complexity of the model increases, including more kinds of interacting cell types and interorgan communication, the analytical complexity also increases. Further, as studies move further into spatially resolved biology, new technical problems have to be overcome owing to the small number of analytes present in the confines of a single cell or cell compartment. Here, we review the overall goals and solutions made possible by stable isotope tracing and their applications to models of increasing complexity. Finally, we discuss progress and outstanding difficulties in high-resolution spatially resolved tracer-based metabolic studies.
摘要:
稳定同位素分辨代谢组学包括一组关键的技术,可应用于各种系统,从分离的细胞到整个生物体,定义代谢途径的使用和对药物或突变等扰动的反应,以及为通量分析提供依据。由于稳定同位素富集化合物的多样性非常高,使用较新的多路复用方法,新陈代谢的覆盖范围现在非常广泛。然而,随着模型复杂性的增加,包括更多种类的相互作用细胞类型和器官间通讯,分析的复杂性也增加了。Further,随着研究进一步进入空间分辨生物学,由于在单个细胞或细胞区室的范围内存在少量的分析物,因此必须克服新的技术问题。这里,我们回顾了通过稳定同位素追踪实现的总体目标和解决方案,以及它们在日益复杂的模型中的应用。最后,我们讨论了高分辨率空间分辨示踪剂代谢研究的进展和突出困难。