Lipid transport across the blood-brain barrier

在这个项目中，我们的目的是阐明选择性脂类通过血脑屏障(BBB)以及主要的筋膜肌超家族结构域包含蛋白2a(Mfsd2a)在这一过程中的作用。我们想测试的假设是，虽然最初涉及Mfsd2a，但介导选择性脂质输入大脑的作用屏障实际上是一个多阶段的过程，包括局部脂质代谢。该项目将研究参与的主要血脑屏障细胞类型、毛细血管内皮细胞和星形胶质细胞的个体贡献，通过表征它们在脂质吸收、代谢和运输方面的活动。此外，还将研究细胞-细胞-细胞连接在这些过程中的作用。利用小鼠模型，将在不断增加的复杂性水平上(体外和体内)进行补充研究。该项目将采用在脂质生物学和新陈代谢研究中开辟新纪元的新型炔脂示踪剂技术。这些非放射性示踪剂与一整套现代分析方法完全兼容，因此可以以前所未有的方式以最高的空间和分析分辨率对脂质定位和代谢进行相关跟踪。我们将把炔脂示踪与最先进的分析相结合，包括超分辨率显微镜和定量质谱仪(MS)脂组学。得益于这一技术优势，这一提议将对哺乳动物大脑选择性摄取脂肪的复杂过程产生重要的见解。

In this project we aim to elucidate the selective lipid transport across the blood brain barrier (BBB) and the role of the major fascilitator superfamily domain-containing protein 2a (Mfsd2a) for this process. We want to test the hypothesis that, while initially involving Mfsd2a, the acting barrier that mediates selective lipid import into the brain in fact is a multi-stage process and includes local lipid metabolism. The project will study the individual contribution of the participating major cell types of the BBB, capillary endothelial cells and astrocytes, by characterizing their activities in lipid uptake, metabolism and trafficking. Also the role of cell-cell-connectivity for these processes will be investigated. Using the mouse model, complementary research at increasing complexity levels (in vitro and in vivo) will be conducted. The project will employ the novel alkyne lipid tracer technology that has opened a new era in lipid biology and metabolism research. These non-radioactive tracers are fully compatible with the suit of modern analytical methods and therefore allow for correlative tracking of lipid localization and metabolism at the highest spatial and analytical resolution in an unprecedented manner. We will combine alkyne lipid tracing with state-of-the-art analytics, including super-resolution microscopy and quantitative mass spectrometry (MS) lipidomics. Benefitting from this technological edge this proposal will yield important insights into the complex process of selective lipid uptake into the mammalian brain.