Department of Biomedicine, Aarhus University, Aarhus C, 8000, Denmark.
Laboratory of Neurological Infection and Immunity, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, 59840, USA.
Fluids Barriers CNS. 2024 Oct 14;21(1):80. doi: 10.1186/s12987-024-00585-x.
Receptor mediated transport of therapeutic antibodies through the blood-brain barrier (BBB) give promise for drug delivery to alleviate brain diseases. We developed a low-cost method to obtain nanoscale localization data of putative cargo receptors. We combine existing ex vivo isolation methods with expansion microscopy (ExM) to analyze receptor localizations in brain microcapillaries. Using this approach, we show how to analyze receptor localizations in endothelial cells of brain microcapillaries in relation to the abluminal marker collagen IV. By choosing the thinnest capillaries, microcapillaries for analysis, we ensure the validity of collagen IV as an abluminal marker. With this tool, we confirm transferrin receptors as well as sortilin to be both luminally and abluminally localized. Furthermore, we identify basigin to be an abluminal receptor. Our methodology can be adapted to analyze different types of isolated brain capillaries and we anticipate that this approach will be very useful for the research community to gain new insight into cargo receptor trafficking in the slim brain endothelial cells to elucidate novel paths for future drug design.
通过血脑屏障（BBB）的受体介导运输治疗性抗体有望实现药物递送，以缓解脑部疾病。我们开发了一种低成本的方法来获得潜在货物受体的纳米级定位数据。我们将现有的离体分离方法与扩展显微镜（ExM）相结合，以分析脑微血管内皮细胞中的受体定位。通过使用这种方法，我们展示了如何分析与基底膜标志物胶原 IV 相关的脑微血管内皮细胞中的受体定位。通过选择最薄的毛细血管作为分析用的微血管，我们确保了胶原 IV 作为基底膜标志物的有效性。使用该工具，我们证实转铁蛋白受体和分选蛋白既是腔侧也是基底侧定位的。此外，我们还确定 basigin 是基底侧受体。我们的方法可以适应分析不同类型的分离脑毛细血管，我们预计这种方法将对研究界非常有用，有助于深入了解货物受体在纤细的脑内皮细胞中的运输，为未来的药物设计阐明新的途径。
