Laser induced wounding of the plasma membrane and methods to study the repair process

Jiménez, Ana Joaquina; Maiuri, Paolo; Lafaurie-Janvore, Julie; Perez, Franck; Piel, Matthieu

doi:10.1016/bs.mcb.2014.11.007

Cited by 7 publications

(4 citation statements)

References 32 publications

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“…Previous studies that analyzed in parallel mechanical- and CDC-pore-induced-wounding concluded that both types of damage triggered a similar repair response (Idone et al, 2008; Corrotte et al, 2013). However, other studies, presented evidence that the size of the wound, for example, affected the mechanism of plasma membrane repair (Jimenez et al, 2015). Therefore, using the CDC-pore-induced membrane damage for the general study of membrane repair is a convenient model, but we cannot rule out that under different damage conditions, there may be other repair mechanisms that are also recruited in response to compromised membrane integrity.…”

Section: Discussionmentioning

confidence: 99%

“…However, controlling the size of the membrane lesions by these approaches is challenging and cells in the population may not be uniformly damaged. Alternatively, micro-needle insertion and laser-induced perforation make a site-specific wound of desired size that can be more precisely controlled and therefore, the reproducibility of the wound is high (Steinhardt et al, 1994; Terasaki et al, 1997; Jimenez et al, 2015). However, these methods work on a single cell and are not amenable to high-throughput screening.…”

Section: Introduction and Limitations Of Current Modelsmentioning

confidence: 99%

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High-Throughput Microplate-Based Assay to Monitor Plasma Membrane Wounding and Repair

Pathak-Sharma

Zhang

Lam

et al. 2017

Front. Cell. Infect. Microbiol.

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The plasma membrane of mammalian cells is susceptible to disruption by mechanical and biochemical damages that frequently occur within tissues. Therefore, efficient and rapid repair of the plasma membrane is essential for maintaining cellular homeostasis and survival. Excessive damage of the plasma membrane and defects in its repair are associated with pathological conditions such as infections, muscular dystrophy, heart failure, diabetes, and lung and neurodegenerative diseases. The molecular events that remodel the plasma membrane during its repair remain poorly understood. In the present work, we report the development of a quantitative high-throughput assay that monitors the efficiency of the plasma membrane repair in real time using a sensitive microplate reader. In this assay, the plasma membrane of living cells is perforated by the bacterial pore-forming toxin listeriolysin O and the integrity and recovery of the membrane are monitored at 37°C by measuring the fluorescence intensity of the membrane impermeant dye propidium iodide. We demonstrate that listeriolysin O causes dose-dependent plasma membrane wounding and activation of the cell repair machinery. This assay was successfully applied to cell types from different origins including epithelial and muscle cells. In conclusion, this high-throughput assay provides a novel opportunity for the discovery of membrane repair effectors and the development of new therapeutic compounds that could target membrane repair in various pathological processes, from degenerative to infectious diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Introduction and Limitations Of Current Modelsmentioning

confidence: 99%

High-Throughput Microplate-Based Assay to Monitor Plasma Membrane Wounding and Repair

Pathak-Sharma

Zhang

Lam

et al. 2017

Front. Cell. Infect. Microbiol.

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“…Here, a single and localized wound can be induced at the plasma membrane of cultured cells and repair can be followed using fast time-lapse imaging [13,41,42]. The main advantages include user's control over the extent of damage by adjusting the laser power and the ability to assess cellular repair kinetic and monitor the action of fluorescently-tagged proteins involved in repair (Figure 4) [43]. A possible drawback is that membrane damage triggered by UV-ablation laser induces local high temperatures at the injured membrane, which can potentially affect membrane proteins and lipids and create thermally-induced diffusion and denaturation artifacts.…”

Section: Approaches To Inflict Damage To the Plasma Membranementioning

confidence: 99%

Interdisciplinary Synergy to Reveal Mechanisms of Annexin-Mediated Plasma Membrane Shaping and Repair

Bendix

Simonsen

Florentsen

et al. 2020

Cells

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The plasma membrane surrounds every single cell and essentially shapes cell life by separating the interior from the external environment. Thus, maintenance of cell membrane integrity is essential to prevent death caused by disruption of the plasma membrane. To counteract plasma membrane injuries, eukaryotic cells have developed efficient repair tools that depend on Ca 2+ -and phospholipid-binding annexin proteins. Upon membrane damage, annexin family members are activated by a Ca 2+ influx, enabling them to quickly bind at the damaged membrane and facilitate wound healing. Our recent studies, based on interdisciplinary research synergy across molecular cell biology, experimental membrane physics, and computational simulations show that annexins have additional biophysical functions in the repair response besides enabling membrane fusion. Annexins possess different membrane-shaping properties, allowing for a tailored response that involves rapid bending, constriction, and fusion of membrane edges for resealing. Moreover, some annexins have high affinity for highly curved membranes that appear at free edges near rupture sites, a property that might accelerate their recruitment for rapid repair. Here, we discuss the mechanisms of annexin-mediated membrane shaping and curvature sensing in the light of our interdisciplinary approach to study plasma membrane repair.

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“…Upon injury, ESCRT complexes promptly accumulate at the wound site, protrude the wounded membrane as a bud or bleb, and subsequently cut it off to release extracellular vesicles. This ESCRT-mediated abscission of the wounded membrane appears to limit smaller-sized wounds (<100 nm in diameter) [145,146].…”

Section: Escrt Complexesmentioning

confidence: 99%

Wound Repair of the Cell Membrane: Lessons from Dictyostelium Cells

Yumura

2024

Cells

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The cell membrane is frequently subjected to damage, either through physical or chemical means. The swift restoration of the cell membrane’s integrity is crucial to prevent the leakage of intracellular materials and the uncontrolled influx of extracellular ions. Consequently, wound repair plays a vital role in cell survival, akin to the importance of DNA repair. The mechanisms involved in wound repair encompass a series of events, including ion influx, membrane patch formation, endocytosis, exocytosis, recruitment of the actin cytoskeleton, and the elimination of damaged membrane sections. Despite the absence of a universally accepted general model, diverse molecular models have been proposed for wound repair in different organisms. Traditional wound methods not only damage the cell membrane but also impact intracellular structures, including the underlying cortical actin networks, microtubules, and organelles. In contrast, the more recent improved laserporation selectively targets the cell membrane. Studies on Dictyostelium cells utilizing this method have introduced a novel perspective on the wound repair mechanism. This review commences by detailing methods for inducing wounds and subsequently reviews recent developments in the field.

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Laser induced wounding of the plasma membrane and methods to study the repair process

Cited by 7 publications

References 32 publications

High-Throughput Microplate-Based Assay to Monitor Plasma Membrane Wounding and Repair

High-Throughput Microplate-Based Assay to Monitor Plasma Membrane Wounding and Repair

Interdisciplinary Synergy to Reveal Mechanisms of Annexin-Mediated Plasma Membrane Shaping and Repair

Wound Repair of the Cell Membrane: Lessons from Dictyostelium Cells

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