评估内吞运输和 Ocrl 功能在斑马鱼神经上皮发育中的作用。
Assessment of endocytic traffic and Ocrl function in the developing zebrafish neuroepithelium.
机构信息
School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, The Michael Smith Building, Oxford Road, Manchester M13 9PT, UK.
出版信息
J Cell Sci. 2022 Sep 15;135(18). doi: 10.1242/jcs.260339. Epub 2022 Sep 20.
Endocytosis allows cells to internalise a wide range of molecules from their environment and to maintain their plasma membrane composition. It is vital during development and for maintenance of tissue homeostasis. The ability to visualise endocytosis in vivo requires suitable assays to monitor the process. Here, we describe imaging-based assays to visualise endocytosis in the neuroepithelium of living zebrafish embryos. Injection of fluorescent tracers into the brain ventricles followed by live imaging was used to study fluid-phase or receptor-mediated endocytosis, for which we used receptor-associated protein (RAP, encoded by Lrpap1) as a ligand for low-density lipoprotein receptor-related protein (LRP) receptors. Using dual-colour imaging combined with expression of endocytic markers, it is possible to track the progression of endocytosed tracers and to monitor trafficking dynamics. Using these assays, we reveal a role for the Lowe syndrome protein Ocrl in endocytic trafficking within the neuroepithelium. We also found that the RAP-binding receptor Lrp2 (encoded by lrp2a) appears to contribute only partially to neuroepithelial RAP endocytosis. Altogether, our results provide a basis to track endocytosis within the neuroepithelium in vivo and support a role for Ocrl in this process. This article has an associated First Person interview with the first author of the paper.
内吞作用使细胞能够从其环境中内化各种分子,并维持其质膜组成。它在发育过程中和组织稳态的维持中至关重要。要在体内可视化内吞作用,需要合适的测定法来监测该过程。在这里,我们描述了基于成像的测定法,以可视化活体斑马鱼胚胎神经上皮中的内吞作用。通过向脑室内注射荧光示踪剂并进行活体成像来研究流体相或受体介导的内吞作用,我们将受体相关蛋白(RAP,由 Lrpap1 编码)用作低密度脂蛋白受体相关蛋白(LRP)受体的配体。使用双色成像结合内吞标记物的表达,可以跟踪内吞示踪剂的进展并监测运输动力学。使用这些测定法,我们揭示了 Lowe 综合征蛋白 Ocrl 在神经上皮内吞运输中的作用。我们还发现,RAP 结合受体 Lrp2(由 lrp2a 编码)似乎仅部分参与神经上皮 RAP 内吞作用。总之,我们的结果为在体内追踪神经上皮内的内吞作用提供了基础,并支持 Ocrl 在该过程中的作用。本文附有该论文第一作者的第一人称采访。
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