Baltes Carsten Alexander, Nolle Friederike, Kaiser Kathi Michèle, Gjana Erbara, Sander Kristin, Jacobs Karin, Hawkins Rhoda Joy, Lautenschläger Franziska
Experimental Physics, Saarland University, Saarbrücken, Germany.
Department of Electrical Engineering, Trier University of Applied Science, Schneidershof, Trier, Germany.
PLoS One. 2025 Sep 5;20(9):e0330071. doi: 10.1371/journal.pone.0330071. eCollection 2025.
Cellular adhesion and contractility are essential for cell movement. In this study, we investigated the effects of actin stabilization on adhesion properties, contractility, and cell migration. For this, we used the recently synthesized actin stabilizer miuraenamide A (MiuA), which has been discussed as a more reliable alternative to the otherwise commonly used actin stabilizer jasplakinolide. We investigated the number and size of focal adhesions in RPE-1 cells and used single-cell force spectroscopy to evaluate the adhesion properties of those cells after MiuA treatment. We showed that MiuA increases the number of focal adhesions while decreasing their size and reduces adhesion energy and force. Additionally, we investigated its effects on the contractility of RPE-1 cells by measuring their contractile energy using pattern-based contractility screening (PaCS). We found no significant change in contractility after MiuA treatment. Finally, we confined RPE-1 cells in PDMS microchannels and analyzed their migration after treatment with MiuA, showing that neither their speed nor their persistence is affected by MiuA. To check that these effects are not specific to RPE-1 cells, we also analyzed the effects of MiuA treatment in MEF cells and neutrophils. Both MEF cells and neutrophils showed the same results as the RPE-1 cells. Our measurements indicate that, although altering focal adhesions significantly reduces adhesion, it does not impact cell contractility. This finding also clarifies why amoeboid migration, which operates independently of adhesion, remains unaffected. Additionally, it explains the previously observed reduction in mesenchymal migration, which relies on adhesion-based mechanisms.
细胞黏附和收缩性对于细胞运动至关重要。在本研究中，我们调查了肌动蛋白稳定化对黏附特性、收缩性和细胞迁移的影响。为此，我们使用了最近合成的肌动蛋白稳定剂三村酰胺A（MiuA），它被认为是比常用的肌动蛋白稳定剂茉莉酮酸内酯更可靠的替代品。我们研究了RPE-1细胞中黏着斑的数量和大小，并使用单细胞力谱来评估MiuA处理后这些细胞的黏附特性。我们发现MiuA增加了黏着斑的数量，同时减小了其大小，并降低了黏附能量和力。此外，我们通过基于模式的收缩性筛选（PaCS）测量RPE-1细胞的收缩能量，研究了MiuA对其收缩性的影响。我们发现MiuA处理后收缩性没有显著变化。最后，我们将RPE-1细胞限制在聚二甲基硅氧烷（PDMS）微通道中，并分析了MiuA处理后的细胞迁移情况，结果表明MiuA既不影响细胞速度也不影响其持续性。为了验证这些效应并非RPE-1细胞所特有，我们还分析了MiuA处理对MEF细胞和中性粒细胞的影响。MEF细胞和中性粒细胞均显示出与RPE-1细胞相同的结果。我们的测量表明，尽管改变黏着斑会显著降低黏附，但并不影响细胞收缩性。这一发现也阐明了为什么独立于黏附的阿米巴样迁移不受影响。此外，它解释了之前观察到的依赖黏附机制的间充质迁移减少的现象。
