Surface morphology live-cell imaging reveals how macropinocytosis inhibitors affect membrane dynamics

Surface morphology live-cell imaging reveals how macropinocytosis inhibitors affect membrane dynamics
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表面形态活细胞成像揭示巨胞饮抑制剂如何影响膜动力学

DOI:
10.1016/j.electacta.2022.141783
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发表时间:
2023
影响因子:
6.6
通讯作者:
Shiku Hitoshi
Shiku Hitoshi
中科院分区:
材料科学2区
文献类型:
--
作者:
Ida Hiroki;Taira Noriko;Azuma Koichi;Kumatani Akichika;Akishiba Misao;Futaki Shiroh;Takahashi Yasufumi;Shiku Hitoshi

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巨胞饮作用是一种独特的内吞途径,涉及动态膜扰动,并且在营养吸收以外的各种细胞功能中很重要。由于其影响,巨胞饮作用已成为各种研究的主题,并且巨胞饮抑制剂已被广泛使用。然而,抑制剂表现出脱靶效应,并且对巨胞饮作用的特异性可能较低,从而影响各种细胞机制,对纳米级膜动力学的影响仍然难以捉摸。在这项研究中,我们利用活细胞的扫描离子电导显微成像揭示了五种不同的抑制剂在治疗前后如何影响膜动力学。我们的结果表明,细胞松弛素和叠氮化钠对水平皱褶运动表现出相似的减缓作用,尽管它们的抑制机制不同。相反,与沿细胞膜方向的运动相比,褶边变形和伸长受到的抑制较弱。这些结果表明还涉及肌动蛋白聚合以外的机制。经过 5-(N-乙基-N-异丙基)-阿米洛利处理后,褶皱消失,细胞表面变得光滑。在存在渥曼青霉素的情况下,我们观察到活跃的褶边,但没有观察到杯状结构。这些发现和扫描离子电导显微镜活细胞成像可能有助于更好地了解抑制剂的作用。
Macropinocytosis is a unique endocytic pathway that involves dynamic membrane ruffling and is important in various cellular functions beyond nutrient uptake. Due to its impact, macropinocytosis has been the subject of various studies and macropinocytosis inhibitors have been widely used. However, inhibitors exhibit offtarget effects and could be less specific for macropinocytosis, thereby affecting various cellular mechanisms with effects on nanoscale membrane dynamics that remain elusive. In this study, we revealed how five different inhibitors affect membrane dynamics before and after treatment using scanning ion conductance microscopic imaging on living cells. Our results indicated that cytochalasins and sodium azide exhibited similar slowing effects on horizontal ruffle movements, even though their inhibitory mechanisms are different. In contrast, ruffle deformation and elongation were less strongly inhibited compared to the movements along cell membrane direction. These results suggested the involvement of mechanisms beyond actin polymerization. The ruffles disappeared and the cell surface smoothened upon the 5-(N-ethyl-N-isopropyl)-Amiloride treatment. In the presence of wortmannin, we observed active ruffles but no cup-like structures. These findings and scanning ion conductance microscopy live-cell imaging would potentially contribute to a better understanding of inhibitor effects.
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影响因子: 5.3
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期刊: FEBS Letters
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DOI: 10.1021/acsnano.1c05202
发表时间: 2021-11-23
期刊: ACS nano
影响因子: 17.1
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影响因子: 3.8
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