Shape Analysis of Biomimetic and Plasma Membrane Vesicles

IF 3.1 Q2 CHEMISTRY, MULTIDISCIPLINARY ChemSystemsChem Pub Date : 2024-10-23 DOI:10.1002/syst.202400052
Rajni Kudawla, Harshmeet Kaur, Tanmay Pandey, Tripta Bhatia
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Abstract

Giant membrane vesicles (GUVs) and Giant plasma membrane vesicles (GPMVs) are used as models to study membrane properties. We conducted a comparative study to examine how reducing the volume of vesicles with different lipid compositions, solution symmetries, solution asymmetries, and membrane charges affects their morphology. We used three-dimensional visualization techniques to study the shape of the vesicles. Although the vesicles may not be perfectly spherical, they exhibit some fluctuations in their shape. To understand these variations, we used confocal image stacks for visualization. Our experimental observations show that the membrane′s charge influences the deflation of the GUVs in the presence of trans-bilayer sugar asymmetries. The lipid bilayers of our GUVs have a uniform distribution of lipids in both leaflets, indicating no asymmetry in lipid composition. We induce trans-bilayer asymmetries by exposing each leaflet of the bilayer to different solution compositions. We also estimated and compared the deformation of GPMV extracted from HEK-293 cells with trans-bilayer buffer asymmetries and inherent leaflet compositional asymmetry with biomimetic membranes.

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仿生囊泡与质膜囊泡的形态分析
以巨膜囊泡(Giant membrane vesicles, GUVs)和巨质膜囊泡(Giant plasma membrane vesicles, GPMVs)为模型研究膜的性质。我们进行了一项比较研究,以研究减少具有不同脂质组成、溶液对称性、溶液不对称性和膜电荷的囊泡的体积如何影响它们的形态。我们使用三维可视化技术来研究囊泡的形状。虽然囊泡可能不是完美的球形,但它们在形状上表现出一些波动。为了理解这些变化,我们使用共聚焦图像堆栈进行可视化。我们的实验观察表明,在跨双层糖不对称存在的情况下,膜的电荷影响了guv的收缩。我们的guv的脂质双层在两个小叶中具有均匀的脂质分布,表明脂质组成不对称。我们通过将双层的每个小叶暴露于不同的溶液组成来诱导跨双层不对称。我们还估计并比较了从HEK-293细胞中提取的具有跨双层缓冲不对称和具有仿生膜的固有小叶组成不对称的GPMV的变形。
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