Observing GLUT4 Translocation in Live L6 Cells Using Quantum Dots

Qu, Feng; Chen, Zubin; Wang, Xiaoxuan; Meng, Lingfeng; Wu, Zheng-Xing; Qu, Ang

doi:10.3390/s110202077

Cited by 3 publications

(4 citation statements)

References 39 publications

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“…[66] They described the relationship between the movements of the QD-kinesin constructs and the dynamics of the cytoskeleton.I nas tudy by Qu et al, [67] QD conjugated to GLUT4 (glucoset ransporter) was internalized into cells by receptor-mediated endocytosis. Despite the difficultieso fQ Dd elivery into the cell, many studies have reported successful intracellular single-molecule QDmediated tracking.…”

Section: Single-molecule Tracking In Living Cellsmentioning

confidence: 99%

“…Courty et al were the first to use QDs for intracellular tracking, in as tudy on the motions of individual kinesin motors in live cells. [66] They described the relationship between the movements of the QD-kinesin constructs and the dynamics of the cytoskeleton.I nas tudy by Qu et al, [67] QD conjugated to GLUT4 (glucoset ransporter) was internalized into cells by receptor-mediated endocytosis. QDs weree asy to observe for al ongert ime than for enhanced GFP,w hich suffers from rapid photobleaching and relativelyw eak fluorescence against ab ackground of strong cellular autofluorescence.…”

Section: Single-molecule Tracking In Living Cellsmentioning

confidence: 99%

“…QDs weree asy to observe for al ongert ime than for enhanced GFP,w hich suffers from rapid photobleaching and relativelyw eak fluorescence against ab ackground of strong cellular autofluorescence. [67] All stages of the in vivo delivery of QD-anti-HER2 conjugates were tracked in real-time: vesselc irculation, extravasation, movement into the extracellular region, bindingt oH ER2 on the cell membrane, and movement from the cell membrane to and into the perinuclear region after endocytosis. [68] Remarkably,Q Dd elivery through the nuclear pore complexh as been observed, by conjugating QDs to moleculest hat penetrate the nuclear membrane, including importins [69] andm RNA.…”

Section: Single-molecule Tracking In Living Cellsmentioning

confidence: 99%

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Quantum Dot‐Based Nanotools for Bioimaging, Diagnostics, and Drug Delivery

2016

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Quantum dots (QDs) are highly fluorescent nanocrystals with advanced photophysical and spectral properties: high brightness and stability against photobleaching accompanied by broad excitation and narrow emission spectra. Water-soluble QDs functionalized with biomolecules, such as proteins, peptides, antibodies, and drugs, are used for biomedical applications. The advantages of QD-based approaches to immuno-histochemical analysis, single-molecule tracking, and in vivo imaging (over traditional methods with organic dyes and fluorescent proteins) are explained. The unique spectral properties of QDs offer opportunities for designing systems for multiplexed analysis by multicolor imaging for the simultaneous detection of multiple targets. Conjugation of drug molecules with QDs or their incorporation into QD-based drug-delivery particles makes it possible to monitor real-time drug tracking and carry out image-guided therapy. Because of the tunability of their photophysical properties, QDs emitting in the near-infrared have become an attractive tool for deep-tissue mono- and multiphoton in vivo imaging. We review recent achievements in QD applications for bioimaging, targeting, and drug delivery, as well as challenges related to their toxicity and non-biodegradability. Key and perspectives for further development of advanced QD-based nanotools are addressed.

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Section: Single-molecule Tracking In Living Cellsmentioning

confidence: 99%

Section: Single-molecule Tracking In Living Cellsmentioning

confidence: 99%

Section: Single-molecule Tracking In Living Cellsmentioning

confidence: 99%

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Quantum Dot‐Based Nanotools for Bioimaging, Diagnostics, and Drug Delivery

2016

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“…Qu et al [87] have applied quantum dots (QDs), which are more photostable and brighter, targeted to GLUT4myc to measure the translocation of GLUT4 in live L6 cells by long-term single particle tracking.…”

Section: Tirf Microscopymentioning

confidence: 99%

Fluorescence microscopy-based quantitation of GLUT4 translocation

Heckmann

Klanert²,

Sandner

et al. 2022

Methods Appl. Fluoresc.

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Postprandial insulin-stimulated glucose uptake into target tissue is crucial for the maintenance of normal blood glucose homeostasis. This step is rate-limited by the number of facilitative glucose transporters type 4 (GLUT4) present in the plasma membrane. Since insulin resistance and impaired GLUT4 translocation are associated with the development of metabolic disorders such as type 2 diabetes, this transporter has become an important target of antidiabetic drug research. The application of screening approaches that are based on the analysis of GLUT4 translocation to the plasma membrane to identify substances with insulinomimetic properties has gained global research interest in recent years. Here, we review methods that have been implemented to quantitate the translocation of GLUT4 to the plasma membrane. These methods can be broadly divided into two sections: microscopy-based technologies (e.g., immunoelectron, confocal or total internal reflection fluorescence microscopy) and biochemical and spectrometric approaches (e.g., membrane fractionation, photoaffinity labeling or flow cytometry). In this review, we discuss the most relevant approaches applied to GLUT4 thus far, highlighting the advantages and disadvantages of these approaches, and we provide a critical discussion and outlook into new methodological opportunities.

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