MemBright: A Family of Fluorescent Membrane Probes for Advanced Cellular Imaging and Neuroscience

Collot, Mayeul; Ashokkumar, Pichandi; Anton, Halina; Boutant, Emmanuel; Faklaris, Orestis; Galli, Thierry; Mély, Yves; Danglot, Lydia; Klymchenko, Andrey S.

doi:10.1016/j.chembiol.2019.01.009

Cited by 166 publications

(190 citation statements)

References 78 publications

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“…On the other hand, a more hydrophobic SQ-535 is probably partially aggregated in PBS, which leads to slower binding kinetics, similarly to previously reported MemBright probes. 8 The aggregation of SQ-535 in PBS is in line with the strong blue shift in absorbance compared to organic solvents, which are much less pronounced in case of FM1-43 and SP-468. Extending the πsystem from SP-468 to SQ-535 led to a 65-67 nm red shift in both absorption and emission spectra of dyes in DOPC liposomes ( Figure 2A).…”

Section: Spectroscopic Studiesmentioning

confidence: 77%

“…From these two observations, numerous efforts have been made to develop fluorescent PM probes 4,5,6,7 including from our group. [8][9][10][11][12][13][14][15] Among them, FM dyes (named after Fei Mao 16 ), which are styryl dyes introduced by Betz and colleagues in the early 90s' as synaptic vesicle markers, 16 rapidly became among the most famous and used commercially available PM probes. 17,18 These amphiphilic dyes were further developed by the group of Loew to give rise to PM voltage sensitive probes 19,20 and membrane lipid domains probes.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging

Collot

Boutant

Fam

et al. 2019

Preprint

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The plasma membrane (PM) plays a major role in many biological processes; therefore its proper fluorescence staining is required in bioimaging. Among the commercially available PM probes, styryl dye FM1-43 is one of the most widely used. In this work, we demonstrated that fine chemical modifications of FM1-43 can dramatically improve the PM staining. The newly developed probes, SP-468 and SQ-535 were found to display enhanced photophysical properties (reduced crosstalk, higher brightness, improved photostability) and unlike FM1-43, provided excellent and immediate PM staining in 5 different mammalian cell lines including neurons (primary culture and tissue imaging). Additionally, we showed that the new probes displayed differences in their internalization pathways compared to their parent FM1-43. Finally, we demonstrated that the modifications made to FM1-43 did not impair the ability of the new probes to stain the PM of plant cells. Overall, this work presents new useful probes for PM imaging in cells and tissues and provides insights on the molecular design of new PM targeting molecules.

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Section: Spectroscopic Studiesmentioning

confidence: 77%

Section: Introductionmentioning

confidence: 99%

Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging

Collot

Boutant

Fam

et al. 2019

Preprint

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“…The ER protein calnexin was present in AB and MV but absent in sEV in line with enrichment in vesicles of endosomal origin in the later. Staining with the lipid probe MemBright, recently developed by Collot and colleagues 46,47 , showed a heterogeneous population of round-shaped vesicles in the AB and MV fractions (Fig.2C). Cryo-electron microscopy images of MV and sEV clearly ascertained the presence of a lipid bilayer membrane surrounding the vesicles (Fig.2D, Suppl.Fig.2).…”

Section: Resultsmentioning

confidence: 90%

Molecular and functional diversity of distinct subpopulations of extracellular vesicles from stressed pancreatic beta cells: implications for autoimmunity

Giri

Beaurepaire

Jégou

et al. 2020

Preprint

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Beta cell failure and apoptosis following islet inflammation have been associated with autoimmune type 1 diabetes pathogenesis. As conveyors of biological active material, extracellular vesicles (EV) act as mediators in communication with immune effectors fostering the idea that EV from inflamed beta cells may contribute to autoimmunity. Evidence accumulates that beta exosomes promote diabetogenic responses, but relative contributions of larger vesicles as well as variations in the composition of the beta cell's vesiculome due to environmental changes have not been explored yet. Here, we made side-by-side comparisons of the phenotype and function of apoptotic bodies (AB), microvesicles (MV) and small EV (sEV) isolated from an equal amount of MIN6 beta cells exposed to inflammatory, hypoxic or genotoxic stressors. Under normal conditions, large vesicles represent 93% of the volume, but only 2% of the number of the vesicles. Our data reveal a consistently higher release of AB and sEV and to a lesser extent of MV, exclusively under inflammatory conditions commensurate with a 4-fold increase in the total volume of the vesiculome and enhanced export of immune-stimulatory material including the autoantigen insulin, microRNA, and cytokines. Whilst inflammation does not change the concentration of insulin inside the EV, specific Toll-like receptor-binding microRNA sequences preferentially partition into sEV. Exposure to inflammatory stress engenders drastic increases in the expression of monocyte chemoattractant protein 1 in all EV and of interleukin-27 solely in AB suggesting selective sorting towards EV subspecies. Functional in vitro assays in mouse dendritic cells and macrophages reveal further differences in the aptitude of EV to modulate expression of cytokines and maturation markers. These findings highlight the different quantitative and qualitative imprints of environmental changes in subpopulations of beta EV that may contribute to the spread of inflammation and sustained immune cell recruitment at the inception of the (auto-) immune response.

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“…MemBright 640nm dye (Collot et al, 2019) in buffer B for 15min at room temperature. Coverslips were mounted in Vectashield and sealed with nail polish.…”

Section: Single-molecule (Sm) Rna Fishmentioning

confidence: 99%

Identification of a Nonsense-Mediated Decay pathway at the Endoplasmic Reticulum

Longman

Jackson-Jones

Maslon

et al. 2020

Preprint

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Nonsense-mediated decay (NMD) is a translation-dependent RNA quality control mechanism that occurs in the cytoplasm. However, it is unknown how NMD regulates the stability of RNAs translated at the Endoplasmic Reticulum (ER). Here, we identify a localized NMD pathway dedicated to ER-translated mRNAs. We previously identified NBAS, a component of the Syntaxin 18 complex involved in Golgi-to-ER trafficking, as a novel NMD factor. Here, we show that NBAS fulfils an independent function in NMD. This ER-NMD pathway requires the interaction of NBAS with the core NMD factor UPF1, which is partially localized at the ER in the proximity of the translocon. NBAS and UPF1 coregulate the stability of ER-associated transcripts, in particular those associated with the cellular stress response. We propose a model where NBAS recruits UPF1 to the membrane of the ER and activates an ER-dedicated NMD pathway, thus providing an ER protective function by ensuring quality control of ER-translated mRNAs.Longman et al. 3 Graphical Abstract HIGHLIGHTS• NBAS is an NMD factor that localizes to the membrane of the endoplasmic reticulum (ER)• NBAS has dual, independent, roles in Golgi-to-ER retrograde transport and in ER-NMD• NBAS recruits the core NMD factor UPF1 to the membrane of the ER• The ER-NMD pathway targets for degradation mRNAs that are translated at the ER ER cy top las m

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MemBright: A Family of Fluorescent Membrane Probes for Advanced Cellular Imaging and Neuroscience

Cited by 166 publications

References 78 publications

Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging

Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging

Molecular and functional diversity of distinct subpopulations of extracellular vesicles from stressed pancreatic beta cells: implications for autoimmunity

Identification of a Nonsense-Mediated Decay pathway at the Endoplasmic Reticulum

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