Molecular rotors report on changes in live cell plasma membrane microviscosity upon interaction with beta-amyloid aggregates

Kubánková, Markéta; López‐Duarte, Ismael; Kiryushko, Darya; Kuimova, Marina K.

doi:10.1039/c8sm01633j

Cited by 34 publications

(44 citation statements)

References 90 publications

(122 reference statements)

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“…GP ranges between +1 and −1, with higher values representing higher lipid order 27,29 . To further interrogate the local PM viscosity, GPMVs were stained with a Bodipy-based molecular rotor 30,31 . The probe lifetime as explored with fluorescence lifetime imaging microscopy (FLIM) is sensitive to the membrane viscosity.…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2019

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Regulation of plasma membrane curvature and composition governs essential cellular processes. The material property of bending rigidity describes the energetic cost of membrane deformations and depends on the plasma membrane molecular composition. Because of compositional fluctuations and active processes, it is challenging to measure it in intact cells. Here, we study the plasma membrane using giant plasma membrane vesicles (GPMVs), which largely preserve the plasma membrane lipidome and proteome. We show that the bending rigidity of plasma membranes under varied conditions is correlated to readout from environment-sensitive dyes, which are indicative of membrane order and microviscosity. This correlation holds across different cell lines, upon cholesterol depletion or enrichment of the plasma membrane, and variations in cell density. Thus, polarity- and viscosity-sensitive probes represent a promising indicator of membrane mechanical properties. Additionally, our results allow for identifying synthetic membranes with a few well defined lipids as optimal plasma membrane mimetics.

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Section: Resultsmentioning

confidence: 99%

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

et al. 2019

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“…Intramolecular rotation is impeded in high viscosity environments, causing higher quantum yields and longer fluorescence lifetimes [16, 17] . Fluorescence lifetime imaging microscopy (FLIM) of molecular rotors allows viscosity mapping by spatially resolving the fluorescence decays of the molecular rotors [13, 14b,c, 17, 18] . Boron dipyrromethene (BODIPY) molecules have emerged as robust and biocompatible rotors with a wide dynamic range of fluorescence lifetimes, from 100 ps to 6 ns, corresponding to a biologically relevant viscosity range of 1 to 5000 cP (centipoise) [14b,c, 18] …”

Section: Figurementioning

confidence: 99%

“…No notable change in average lifetime was observed upon incubation with CORM‐2 (Figures 5 F–J). Using the fluorescence lifetime‐viscosity calibration of 6⋅CO , the average lifetime of 3.3 ns corresponded to a viscosity of 133 cP, a typical value for lipid‐based intracellular organelles [13, 14b,c, 18] …”

Section: Figurementioning

confidence: 99%

Simultaneous Detection of Carbon Monoxide and Viscosity Changes in Cells

et al. 2020

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A new family of robust, non-toxic, water-compatible ruthenium(II) vinyl probes allows the rapid, selective and sensitive detection of endogenous carbon monoxide (CO) in live mammalian cells under normoxic and hypoxic conditions. Uniquely, these probes incorporate a viscosity-sensitive BODIPY fluorophore that allows the measurement of microscopic viscosity in live cells via fluorescence lifetime imaging microscopy (FLIM) while also monitoring CO levels. This is the first example of a probe that can simultaneously detect CO alongside small viscosity changes in organelles of live cells. Carbon monoxide (CO) has long been associated with its toxicity, however, this colourless and odourless gas also plays a key role in cellular messaging. [1] Its anti-inflammatory, antiproliferative, anti-apoptotic and anti-coagulative properties are now recognised. Intriguingly, under pathophysiological conditions (e.g., inflammation) CO production in cells increases, [2] and the real-time monitoring of these changes could potentially provide diagnostic information. Haemoglobin is required as a substrate for CO production in vivo and the haem oxygenases (HO-1 and HO-2) play a key role in the generation of this gas in mammals. Emerging evidence suggests that the increased generation of HO-derived CO plays a critical role in the resolution of inflammatory processes and alleviation of cardiovascular disorders, [3] which has driven interest in CO-releasing molecules (CORMs) for therapy. [4] A major obstacle is the lack of effective methods to track CO in biological systems in real time. [5] Imaging with emissive probes has emerged as one of the most powerful techniques to detect biologically important molecules. However, designing selective CO probes for the cellular environment is challenging, with its wide range of reactive species and variation in pH. He [6] and Chang [7] pioneered two very different

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“…We then imaged PM viscosity by FLIM using membrane-inserting BODIPY-based molecular rotors as previously described ( 34 , 35 ). It has been demonstrated that BODIPY rotors localize in the acyl chain regions of fluid-phase bilayers ( 36 ) and, upon excitation, can either undergo radiative decay via fluorescence emission or decay via nonradiative pathways, typically involving an intramolecular rotation mechanism.…”

Section: Resultsmentioning

confidence: 99%

Rapid formation of human immunodeficiency virus-like particles

Bednarska

Pelchen–Matthews

Novák

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Understanding the molecular mechanisms involved in the assembly of viruses is essential for discerning how viruses transmit from cell to cell and host to host. Although molecular aspects of assembly have been studied for many viruses, we still have little information about these events in real time. Enveloped viruses such as HIV that assemble at, and bud from, the plasma membrane have been studied in some detail using live cell fluorescence imaging techniques; however, these approaches provide little information about the real-time morphological changes that take place as viral components come together to form individual virus particles. Here we used correlative scanning ion conductance microscopy and fluorescence confocal microscopy to measure the topological changes, together with the recruitment of fluorescently labeled viral proteins such as Gag and Vpr, during the assembly and release of individual HIV virus-like particles (VLPs) from the top, nonadherent surfaces of living cells. We show that 1) labeling of viral proteins with green fluorescent protein affects particle formation, 2) the kinetics of particle assembly on different plasma membrane domains can vary, possibly as a consequence of differences in membrane biophysical properties, and 3) VLPs budding from the top, unimpeded surface of cells can reach full size in 20 s and disappear from the budding site in 0.5 to 3 min from the moment curvature is initially detected, significantly faster than has been previously reported.

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Molecular rotors report on changes in live cell plasma membrane microviscosity upon interaction with beta-amyloid aggregates

Abstract: Viscosity-sensing fluorophores termed molecular rotors were used to investigate the microviscosity of plasma membranes of live cells upon interaction with oligomeric and fibrillar forms of beta-amyloid Aβ(1–42).

Cited by 34 publications

References 90 publications

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

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

Simultaneous Detection of Carbon Monoxide and Viscosity Changes in Cells

Rapid formation of human immunodeficiency virus-like particles

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