细胞单层和三维多细胞聚集体中长春花素张力的比较。

IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Luni Hu,Rick I Cohen,Margarida Barroso,Nada N Boustany
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引用次数: 0

摘要

对表达长春花素张力传感器(VinTS)的中国仓鼠卵巢(CHO-K1)细胞进行共聚焦频域荧光寿命和福斯特共振能量转移(FRET)显微镜观察,以比较长春花素在三维(3D)多细胞聚集体和二维细胞单层中的张力。在二维和三维培养物中,VinTS 的 FRET 效率比 VinTL 低 5-6%(p < 0.05),VinTL 是一种不能结合肌动蛋白或 paxillin 的无尾对照。用 Rho- 相关激酶抑制剂 Y-27632 处理可减轻 VinTS 和 VinTL FRET 效率之间的差异,这表明 VinTS 在二维和三维培养物中都处于紧张状态。然而,与二维单层培养物相比,三维多细胞聚集体中 VinTS 和 VinTL 的 FRET 效率总体下降。VinTS 在二维和三维培养物中的表达呈现出与细胞粘附一致的点状。虽然 paxillin 存在于二维单层培养物中 VinTS 的表达位点,但在三维聚集体中 VinTS 的表达点一般不存在 paxillin。这些结果表明,与二维单层相比,VinTS 在三维聚集体中经历的环境有所改变,这为进一步研究三维组织模型中的分子张力传感器提供了基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of vinculin tension in cellular monolayers and three-dimensional multicellular aggregates.
Confocal frequency-domain fluorescence lifetime and Förster resonance energy transfer (FRET) microscopy of Chinese hamster ovary (CHO-K1) cells expressing the vinculin tension sensor (VinTS) is used to compare vinculin tension in three-dimensional (3D) multicellular aggregates and 2D cellular monolayers. In both 2D and 3D cultures, the FRET efficiency of VinTS is 5-6% lower than that of VinTL (p < 0.05), a tail-less control which cannot bind actin or paxillin. The difference between VinTS and VinTL FRET efficiency can be mitigated by treatment with the Rho-associated kinase inhibitor Y-27632, demonstrating that VinTS is under tension in both 2D and 3D cultures. However, there is an overall decrease in FRET efficiency of both VinTS and VinTL in the 3D multicellular aggregates compared with the 2D monolayers. Expression of VinTS in 2D and 3D cultures exhibits puncta consistent with cellular adhesions. While paxillin is present at the sites of VinTS expression in the 2D monolayers, it is generally absent from VinTS puncta in the 3D aggregates. The results suggest that VinTS experiences a modified environment in 3D aggregates compared with 2D monolayers and provide a basis for further investigation of molecular tension sensors in 3D tissue models.
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来源期刊
Biomedical optics express
Biomedical optics express BIOCHEMICAL RESEARCH METHODS-OPTICS
CiteScore
6.80
自引率
11.80%
发文量
633
审稿时长
1 months
期刊介绍: The journal''s scope encompasses fundamental research, technology development, biomedical studies and clinical applications. BOEx focuses on the leading edge topics in the field, including: Tissue optics and spectroscopy Novel microscopies Optical coherence tomography Diffuse and fluorescence tomography Photoacoustic and multimodal imaging Molecular imaging and therapies Nanophotonic biosensing Optical biophysics/photobiology Microfluidic optical devices Vision research.
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