随机光学重建显微镜(STORM)观察内皮表面糖萼(ESG)成分及超微结构。

IF 1 4区医学 Q4 BIOPHYSICS

Biorheology Pub Date : 2019-01-01 DOI:10.3233/BIR-180204

Jie Fan, Yi Sun, Yifan Xia, John M Tarbell, Bingmei M Fu

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引用次数: 21

摘要

背景:内皮表面糖萼(endothelial surface glycocalyx, ESG)应具有有组织的结构，以在血管功能中发挥不同的作用，如血液流动的机械传感器和跨血管交换的屏障。由于光学和电子显微镜的限制，ESG的超结构直到最近发展的超分辨率光学显微镜STORM才被揭示出来。目的:研究小鼠脑微血管内皮细胞(bEnd3)单层上ESG组分及其组织结构。方法:用抗硫酸肝素(HS)免疫标记ESG，然后用ATTO488偶联山羊抗小鼠IgG免疫标记ESG，用生物素化透明质酸(HA)结合蛋白免疫标记ESG，最后用AF647偶联抗生物素免疫标记ESG。然后由STORM对ESG进行成像。结果:透明质酸是一个长分子编织成一个网络，覆盖内皮腔表面。相反，HS是一个更短的分子，垂直于细胞表面。HA和HS在内皮腔表面部分重叠。我们还量化了内皮表面上、中、下区域HS的长度、直径、方向和密度。结论:我们的研究结果表明HS在机械传感中起主要作用，HA在分子筛中起主要作用。

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Endothelial surface glycocalyx (ESG) components and ultra-structure revealed by stochastic optical reconstruction microscopy (STORM).

Background: In order to play different roles in vascular functions as a mechanosensor to blood flows and as a barrier to transvascular exchange, the endothelial surface glycocalyx (ESG) should have an organized structure. Due to the limitations of optical and electron microscopy, the ultra-structure of ESG has not been revealed until the recent development of super-resolution optical microscopy, STORM.

Objectives: To investigate the ESG components and their organization on bEnd3 (mouse brain microvascular endothelial cells) monolayer.

Methods: ESG was immunolabeled with anti-heparan sulfate (HS), followed by an ATTO488 conjugated goat anti-mouse IgG, and with biotinylated hyaluronic acid (HA) binding protein, followed by an AF647 conjugated anti-biotin. The ESG was then imaged by the STORM.

Results: HA is a long molecule weaving into a network which covers the endothelial luminal surface. In contrast, HS is a shorter molecule, perpendicular to the cell surface. HA and HS are partially overlapped with each other at the endothelial luminal surface. We also quantified the length, diameter, orientation, and density of HS at the top, middle and bottom regions of the endothelial surface.

Conclusions: Our results suggest that HS plays a major role in mechanosensing and HA plays a major role in the molecular sieve.

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来源期刊

Biorheology 医学-工程：生物医学

CiteScore

2.00

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Biorheology is an international interdisciplinary journal that publishes research on the deformation and flow properties of biological systems or materials. It is the aim of the editors and publishers of Biorheology to bring together contributions from those working in various fields of biorheological research from all over the world. A diverse editorial board with broad international representation provides guidance and expertise in wide-ranging applications of rheological methods to biological systems and materials. The scope of papers solicited by Biorheology extends to systems at different levels of organization that have never been studied before, or, if studied previously, have either never been analyzed in terms of their rheological properties or have not been studied from the point of view of the rheological matching between their structural and functional properties. This biorheological approach applies in particular to molecular studies where changes of physical properties and conformation are investigated without reference to how the process actually takes place, how the forces generated are matched to the properties of the structures and environment concerned, proper time scales, or what structures or strength of structures are required.