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全面内源性标记基底膜成分揭示了基质支架内的动态运动。

Comprehensive Endogenous Tagging of Basement Membrane Components Reveals Dynamic Movement within the Matrix Scaffolding.

机构信息

Department of Biology, Duke University, Box 90338, Durham, NC 27708, USA.

Department of Biology, Duke University, Box 90338, Durham, NC 27708, USA; Department of Cell Biology, Duke University, Box 3709, Durham, NC 27710, USA.

出版信息

Dev Cell. 2020 Jul 6;54(1):60-74.e7. doi: 10.1016/j.devcel.2020.05.022. Epub 2020 Jun 24.

Abstract

Basement membranes (BMs) are supramolecular matrices built on laminin and type IV collagen networks that provide structural and signaling support to tissues. BM complexity, however, has hindered an understanding of its formation, dynamics, and regulation. Using genome editing, we tagged 29 BM matrix components and receptors in C. elegans with mNeonGreen. Here, we report a common template that initiates BM formation, which rapidly diversifies during tissue differentiation. Through photobleaching studies, we show that BMs are not static-surprisingly, many matrix proteins move within the laminin and collagen scaffoldings. Finally, quantitative imaging, conditional knockdown, and optical highlighting indicate that papilin, a poorly studied glycoprotein, is the most abundant component in the gonadal BM, where it facilitates type IV collagen removal during BM expansion and tissue growth. Together, this work introduces methods for holistic investigation of BM regulation and reveals that BMs are highly dynamic and capable of rapid change to support tissues.

摘要

基底膜 (BMs) 是构建在层粘连蛋白和 IV 型胶原网络上的超分子基质,为组织提供结构和信号支持。然而,BM 的复杂性阻碍了对其形成、动态和调控的理解。我们使用基因组编辑技术,在 C. elegans 中标记了 29 种 BM 基质成分和受体,并用 mNeonGreen 进行标记。在这里,我们报告了一个启动 BM 形成的通用模板,该模板在组织分化过程中迅速多样化。通过光漂白研究,我们表明 BM 不是静态的——令人惊讶的是,许多基质蛋白在层粘连蛋白和胶原支架内移动。最后,定量成像、条件性敲低和光学突出显示表明,papilin 是一种研究较少的糖蛋白,是性腺 BM 中最丰富的成分,它在 BM 扩张和组织生长过程中促进 IV 型胶原的去除。总之,这项工作介绍了全面研究 BM 调控的方法,并揭示了 BM 具有高度动态性和快速变化的能力,以支持组织。

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