Di-4-ANEPPDHQ, a fluorescent probe for the visualisation of membrane microdomains in living Arabidopsis thaliana cells

Zhao, Xiaoyu; Li, Ruili; Lu, Cunfu; Baluška, Frantǐsek; Wan, Yinglang

doi:10.1016/j.plaphy.2014.12.015

Cited by 31 publications

(19 citation statements)

References 25 publications

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“…Here, we observed changes in lipid order in the endosomal and PM compartments. Quantitative images showed that endosomes have lower GP values than those of the PM in all of the cell types we analyzed, in agree with the early reports ( Frescatada-Rosa et al, 2014 ; Zhao et al, 2015 ). Frescatada-Rosa used the di-4-ANEPPDHQ to quantify the lipid orders in Arabidopsis as well, however, the GP values in this report is much lower than the values we got.…”

Section: Discussionsupporting

confidence: 93%

“…Namely, concentrated membrane microdomains can increase lipid order, resulting in the high GP values. In consideration of the relative high cytotoxity of filipin to plant cells ( Ovečka et al, 2010 ; Boutté et al, 2011 ), we have used di-4-ANEPPDHQ as an optimal probe to visualize and quantify the lipid order in membranes of living plant cells ( Zhao et al, 2015 ). Moreover, di-4-ANEPPDHQ was used for imaging pf polarly growing pollen tubes as well, and it was clearly reported that the apical regions of Nicotiana tabacum, Picea meyeri have significant higher GP values than other regions ( Liu et al, 2009 ; Moscatelli et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

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Mapping of Membrane Lipid Order in Root Apex Zones of Arabidopsis thaliana

Zhao

Zhang

et al. 2015

Front. Plant Sci.

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In this study, we used the fluorescence probe, Di-4-ANEPPDHQ, to map the distribution of membrane lipid order in the apical region of Arabidopsis roots. The generalized polarization (GP) value of Di-4-ANEPPDHQ-stained roots indicated the highest lipid order in the root transition zone (RTZ). The cortical cells have higher lipid order than the epidermal cells in same regions, while the developing root hairs show very prominent cell polarity with high lipid order in apical region. Moreover, the endosomes had lower lipid order than that of the plasma membrane (PM). Brefeldin A (BFA) treatment decreased the lipid order in both the plasma and endosomal membranes of epidermal cells in the RTZ. The lipid order of BFA-induced compartments became higher than that of the PM after BFA treatment in epidermal cells. Meanwhile, the polarly growing tips of root hairs did not show the same behavior. The lipid order of the PM remained unchanged, with higher values than that of the endosomes. This suggests that the lipid ordering in the PM was affected by recycling of endosomal vesicles in epidermal cells of the root apex transition zone but not in the root hairs of Arabidopsis.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Mapping of Membrane Lipid Order in Root Apex Zones of Arabidopsis thaliana

Zhao

Zhang

et al. 2015

Front. Plant Sci.

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“…Proton pump PMA fused to GFP was used as control of membrane integrity after fen treatment ( Figure 1—figure supplement 5B–E ). To obtain further evidence of the enrichment of St REM1.3 into sterol-enriched nanodomains, expected to display a higher degree of order ( Dufourc, 2008 ), we used the environment–sensitive probe di-4-ANEPPDHQ in vivo ( Zhao et al, 2015 ). Figure 1D shows that nanodomains enriched in YFP- St REM1.3 co-localized with highly ordered regions of the PM, in good agreement with the involvement of sterols in St REM1.3 localization.…”

Section: Resultsmentioning

confidence: 99%

Structural basis for plant plasma membrane protein dynamics and organization into functional nanodomains

Gronnier

Crowet

Habenstein

et al. 2017

eLife

131

167

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Plasma Membrane is the primary structure for adjusting to ever changing conditions. PM sub-compartmentalization in domains is thought to orchestrate signaling. Yet, mechanisms governing membrane organization are mostly uncharacterized. The plant-specific REMORINs are proteins regulating hormonal crosstalk and host invasion. REMs are the best-characterized nanodomain markers via an uncharacterized moiety called REMORIN C-terminal Anchor. By coupling biophysical methods, super-resolution microscopy and physiology, we decipher an original mechanism regulating the dynamic and organization of nanodomains. We showed that targeting of REMORIN is independent of the COP-II-dependent secretory pathway and mediated by PI4P and sterol. REM-CA is an unconventional lipid-binding motif that confers nanodomain organization. Analyses of REM-CA mutants by single particle tracking demonstrate that mobility and supramolecular organization are critical for immunity. This study provides a unique mechanistic insight into how the tight control of spatial segregation is critical in the definition of PM domain necessary to support biological function.DOI: http://dx.doi.org/10.7554/eLife.26404.001

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“…It has an emission maximum of 440 nm and 490 nm in gel and liquid crystalline phase membranes, respectively [11]. In comparison, the more recent probe di-4-ANEPPDHQ [12] has a red-shifted spectra with emission maximum at 560 nm or 610 nm in gel or liquid crystalline phase membranes, respectively [13]. The spectral shift is used to calculate the generalized polarization (GP), which is a relative quantitative measure for lipid packing [14,15].…”

Section: Introductionmentioning

confidence: 99%

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

Amaro

Reina

Hof

et al. 2016

Preprint

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Lipid packing is a crucial feature of cellular membranes. Quantitative analysis of membrane lipid packing can be achieved using polarity sensitive probes whose emission spectrum depends on the lipid packing. However, detailed insights into the exact mechanisms that cause the changes in the spectra are necessary to interpret experimental fluorescence emission data correctly. Here, we analysed frequently used polarity sensitive probes, Laurdan and di-4-ANEPPDHQ, to test whether the underlying physical mechanisms of their spectral changes are the same and, thus, whether they report on the same physico-chemical properties of the cell membrane. Steady-state spectra as well as time-resolved emission spectra of the probes in solvents and model membranes revealed that they probe different properties of the lipid membrane. Our findings are important for the application of these dyes in cell biology.

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Di-4-ANEPPDHQ, a fluorescent probe for the visualisation of membrane microdomains in living Arabidopsis thaliana cells

Cited by 31 publications

References 25 publications

Mapping of Membrane Lipid Order in Root Apex Zones of Arabidopsis thaliana

Mapping of Membrane Lipid Order in Root Apex Zones of Arabidopsis thaliana

Structural basis for plant plasma membrane protein dynamics and organization into functional nanodomains

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

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