Characterization of a new class of blue-fluorescent lipid droplet markers for live-cell imaging in plants

Kuntam, Soujanya; Puskás, László G.; Ayaydin, Ferhan

doi:10.1007/s00299-015-1738-4

Cited by 17 publications

(20 citation statements)

References 39 publications

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“…31 In order to complete the available palette of color, blue emitting LDs markers have recently been reported. 37,38,39,40 Although they showed good selectivity and despite an example where two-photon excitation was used, 35 their brightness in imaging are quite limited due to their UV shifted excitation spectra and quite low molar absorption coefficients, thus leading to low excitation efficiency using the common 405 nm laser. Except one example where a near infrared fluorophores served to visualize the most viscous LDs, 41 LDs probes being able to report their environment such as composition or polarity using fluorescence imaging have not yet been reported.…”

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confidence: 99%

Probing Polarity and Heterogeneity of Lipid Droplets in Live Cells Using a Push–Pull Fluorophore

et al. 2018

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Lipid droplets (LDs) are organelles composed of a lipid core surrounded by a phospholipid monolayer. Lately, LDs attracted a considerable attention due to recent studies demonstrating their role in a variety of physiological processes as well as diseases. Herein we synthesized a push-pull molecule named DAF (Dimethyl Aniline Furaldehyde) that possesses a strong positive solvatochromism in emission of 119 nm from toluene to methanol. Its impressive fluorogenic properties from water to oil (2000fold) as well as its high quantum yields (up to 0.97) led us to investigate its ability to sense the distribution of polarity in live cells by fluorescence ratiometric imaging. When added to live cells and excited at 405 nm, DAF immediately and brightly stain lipid droplets using a blue channel (410-500 nm) and cytoplasm in a red channel (500-600 nm). DAF also proved to be compatible with fixation thus allowing 3D imaging of LDs in their cytoplasm environment. Taking advantage of DAF emission in two distinct channels, ratiometric imaging was successfully performed and led to the polarity mapping of the cell unraveling some heterogeneity in polarity within LDs of the same cell.

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confidence: 99%

Probing Polarity and Heterogeneity of Lipid Droplets in Live Cells Using a Push–Pull Fluorophore

et al. 2018

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“…For the Nile red staining, 4 μL of stock solution (500 μg mL -1 dissolved in acetone) was added to 1 mL of 75% glycerol, some droplets were added on a glass slide containing air-dried sections, a coverslip was placed and observed immediately with a LSM 510 META confocal microscope; the fluorophore was excited with a 488 and 514 nm laser lines and emission was filtered with a BP 575–615 IR filter ( Kuntam et al, 2015 ).…”

Section: Methodsmentioning

confidence: 99%

Exploring Cell Wall Composition and Modifications During the Development of the Gynoecium Medial Domain in Arabidopsis

Herrera-Ubaldo

Folter

2018

Front. Plant Sci.

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In Arabidopsis, the gynoecium, the inner whorl of the flower, is the female reproductive part. Many tissues important for fertilization such as the stigma, style, transmitting tract, placenta, ovules, and septum, comprising the medial domain, arise from the carpel margin meristem. During gynoecium development, septum fusion occurs and tissues form continuously to prepare for a successful pollination and fertilization. During gynoecium development, cell wall modifications take place and one of the most important is the formation of the transmitting tract, having a great impact on reproductive competence because it facilitates pollen tube growth and movement through the ovary. In this study, using a combination of classical staining methods, fluorescent dyes, and indirect immunolocalization, we analyzed cell wall composition and modifications accompanying medial domain formation during gynoecium development. We detected coordinated changes in polysaccharide distribution through time, cell wall modifications preceding the formation of the transmitting tract, mucosubstances increase during transmitting tract formation, and a decrease of mannan distribution. Furthermore, we also detected changes in lipid distribution during septum fusion. Proper cell wall composition and modifications are important for postgenital fusion of the carpel (septum fusion) and transmitting tract formation, because these tissues affect plant reproductive competence.

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“…Cytoplasmic LD can be stained in vivo with different dyes. We compared the commonly used Nile red (James et al, 2010;McLachlan et al, 2016) to LD540 (Spandl et al, 2009), and the recently described blue-fluorescent LD dyes for live-cell imaging in plant (Kuntam et al, 2015). We found that LD540 perfectly co-localized with the well characterized LD marker ERG6-RFP in yeast with little or no staining of non-LD structures ( Figure S2).…”

Section: Attspo Can Modulate the Level Of Cytoplasmic Ld In Arabidopsismentioning

confidence: 99%

The multistress‐induced Translocator protein (TSPO) differentially modulates storage lipids metabolism in seeds and seedlings

et al. 2018

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Translocator proteins (TSPO) are conserved membrane proteins extensively studied in mammals, but their function is still unclear. Angiosperm TSPO are transiently induced by abiotic stresses in vegetative tissues. We showed previously that constitutive expression of the Arabidopsis TSPO (AtTSPO) could be detrimental to the cell. Degradation of AtTSPO requires an active autophagy pathway. We show here that genetic modifications of TSPO expression in plant and yeast cells reduce the levels of cytoplasmic lipid droplets (LD). Transgenic Arabidopsis seedlings overexpressing AtTSPO contain less LD as compared with wild type (WT). LD levels were increased in Arabidopsis AtTSPO knockout (KO) seedlings. Deletion of the Schizosaccharomyces pombe TSPO resulted in an increase in LD level in the cell. As compared with the WT, the mutant strain was more sensitive to cerulenin, an inhibitor of fatty acids and sterol biosynthesis. We found that in contrast with seedlings, overexpression of AtTSPO (OE) resulted in an up to 50% increase in seeds fatty acids as compared with WT. A time course experiment revealed that after 4 days of seed imbibition, the levels of triacylglycerol (TAG) was still higher in the OE seeds as compared with WT or KO seeds. However, the de novo synthesis of phospholipids and TAG after 24 h of imbibition was substantially reduced in OE seeds as compared with WT or KO seeds. Our findings support a plant TSPO role in energy homeostasis in a tissue-specific manner, enhancing fatty acids and LD accumulation in mature seeds and limiting LD levels in seedlings.

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Characterization of a new class of blue-fluorescent lipid droplet markers for live-cell imaging in plants

Cited by 17 publications

References 39 publications

Probing Polarity and Heterogeneity of Lipid Droplets in Live Cells Using a Push–Pull Fluorophore

Probing Polarity and Heterogeneity of Lipid Droplets in Live Cells Using a Push–Pull Fluorophore

Exploring Cell Wall Composition and Modifications During the Development of the Gynoecium Medial Domain in Arabidopsis

The multistress‐induced Translocator protein (TSPO) differentially modulates storage lipids metabolism in seeds and seedlings

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