Laurdan and Di-4-ANEPPDHQ probe different properties of the membrane

Amaro, Mariana; Reina, Francesco; Hof, Martin; Eggeling, Christian; Sezgin, Erdinç

doi:10.1101/076752

Cited by 18 publications

(29 citation statements)

References 42 publications

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“…Confocal images were analyzed as previously described (25,26). Briefly, for each experimental condition, ca.…”

Section: Image Processingmentioning

confidence: 99%

“…In order to investigate the effects of EPC3 directly in the PM of living cells, we applied an approach based on the spectroscopic properties of two fluorescent molecules (i.e. Di-4-ANEPPDHQ and Laurdan) which are strongly sensitive to membrane order and lipid packing (25,26). In the context of studies regarding the effect of APLs on lipid bilayers, Laurdan was used to detect and increase in the fluidity of model membranes induced by Miltefosine (39).…”

Section: Epc3 Alters the Fluidity Of The Plasma Membrane Of Living Cellsmentioning

confidence: 99%

“…S1), thus making ROI selections more difficult and less precise, while generally increasing the spread in observed GP values. Second, Di-4-ANEPPDQH and Laurdan are sensitive to different properties of the lipid bilayer (25). While Laurdan is sensitive to lipid packing, the spectroscopic properties of Di-4-ANEPPDQH are influenced by other factors, such as cholesterol content of the membrane or internal electric dipole potential of the bilayer.…”

Section: Epc3 Alters the Fluidity Of The Plasma Membrane Of Living Cellsmentioning

confidence: 99%

“…Second, we quantified the influence of EPC3 on the membrane order for the PM of cancer cell models by using polarity-sensitive fluorescent probes (25). More in detail, we have investigated the spectral properties of two membrane probes (Laurdan and Di-4-ANEPPDHQ) which were shown to be influenced by lipid packing, membrane hydration and composition to different extents (25,26).…”

Section: Introductionmentioning

confidence: 99%

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Effect of Erufosine on Membrane Lipid Order in Breast Cancer Cell Models

Tzoneva

Stoyanova

Petrich

et al. 2020

Preprint

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Alkylphospholipids are a novel class of antineoplastic drugs showing remarkable therapeutic potential. Among them, Erufosine (EPC3) is a promising drug for the treatment of several types of tumors which has not been sufficiently characterized. While EPC3 is supposed to exert its function by interacting with lipid membranes, the exact molecular mechanisms involved are not known yet. In this work, we applied a combination of several fluorescence microscopy techniques (i.e., scanning fluorescence correlation spectroscopy, line-scan fluorescence correlation spectroscopy and generalized polarization imaging) to quantify the effect of EPC3 in biophysical models of the plasma membrane, as well as in cancer cell lines.Our results indicate that EPC3 affects lipid-lipid interactions in cellular membranes by decreasing lipid packing and increasing membrane disorder and fluidity. As a consequence of these alterations in the lateral organization of lipid bilayers, the diffusive dynamics of membrane proteins are also significantly increased. Taken together, these findings suggest that the mechanism of action of EPC3 might be linked to its effects on fundamental biophysical properties of lipid membranes in cancer cells.

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“…Confocal images were analyzed as previously described (25,26). Briefly, for each experimental condition, ca.…”

Section: Image Processingmentioning

confidence: 99%

Section: Epc3 Alters the Fluidity Of The Plasma Membrane Of Living Cellsmentioning

confidence: 99%

Section: Epc3 Alters the Fluidity Of The Plasma Membrane Of Living Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of Erufosine on Membrane Lipid Order in Breast Cancer Cell Models

Tzoneva

Stoyanova

Petrich

et al. 2020

Preprint

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“…Apparently, the artefacts induced by DTT do not exhibit a strong effect on the bending rigidity. We also questioned the effect of PFA (34), which may be potentially inducing (membrane) protein crosslinking, even when used at much lower concertation then in cell fixation protocols. GPMVs were extracted using NEM, and 25 mM PFA was added to the GPMV suspension followed by 1 hour incubation.…”

Section: Influence Of Gpmv Isolation Chemicalsmentioning

confidence: 99%

Mechanical properties of plasma membrane vesicles correlate with lipid order and viscosity and depend on cell density

Steinkühler

Sezgin

Urbančič

et al. 2019

Preprint

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Plasma membranes dynamically respond to external cues and changing environment. Quantitative measurements of these adaptations can elucidate the mechanism that cells exploit to survive, adapt and function. However, cell-based assays are affected by active processes while measurements on synthetic models suffer from compositional limitations.Here, as a model system we employ giant plasma membrane vesicles (GPMVs), which largely preserve the plasma membrane lipidome and proteome. From analysis of fluorescence emission and lifetime of environment-sensitive dyes, and membrane shape fluctuations, we investigate how plasma membrane order, viscosity and bending rigidity are affected by different stimuli such as cell seeding density in three different cell models. Our studies reveal that bending rigidity of plasma membranes vary with lipid order and microviscosity in a highly correlated fashion. Thus, readouts from polarity-and viscosity-sensitive probes represent a promising indicator of membrane mechanical properties. Quantitative analysis of the data allows for comparison to synthetic lipid membranes as plasma membrane mimetics.

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