Quantitative single particle tracking of NGF–receptor complexes: Transport is bidirectional but biased by longer retrograde run lengths

Echarte, Marı́a Mercedes; Bruno, Luciana; Arndt‐Jovin, Donna J.; Jovin, Thomas M.; Pietrasanta, Lı́a I.

doi:10.1016/j.febslet.2007.05.041

Cited by 47 publications

(38 citation statements)

References 50 publications

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“…Biotinylated ligands in combination with streptavidin-conjugated quantum dots (svQDs) provide valuable tools for studying the dynamics of RTKs in living cells, including the characterization of the internalization process and retrograde transport (Lidke et al, 2004;Lidke et al, 2005;Lidke et al, 2007;Cambi et al, 2007;Echarte et al, 2007;Andrews et al, 2009). The combination of the expression of IR-VFPs, binding to the IR of BAC-Ins and its detection with QDs constituted a sensitive monitor of insulin-IR complexes in membranes and on filopodia of HeLa cells.…”

Section: Biotinamido-caproyl Insulin (Bac-ins) and Quantum Dots Bind mentioning

confidence: 99%

Differential endocytosis and signaling dynamics of insulin receptor variants IR-A and IR-B

Giudice

Leskow

Arndt‐Jovin

et al. 2011

Journal of Cell Science

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Insulin signaling comprises a complex cascade of events, playing a key role in the regulation of glucose metabolism and cellular growth. Impaired response to insulin is the hallmark of diabetes, whereas upregulated insulin activity occurs in many cancers. Two splice variants of the insulin receptor (IR) exist in mammals: IR-A, lacking exon 11, and full-length IR-B. Although considerable biochemical data exist on insulin binding and downstream signaling, little is known about the dynamics of the IR itself. We created functional IR transgenes fused with visible fluorescent proteins for use in combination with biotinamido-caproyl insulin and streptavidin quantum dots. Using confocal and structured illumination microscopy, we visualized the endocytosis of both isoforms in living and fixed cells and demonstrated a higher rate of endocytosis of IR-A than IR-B. These differences correlated with higher and sustained activation of IR-A in response to insulin and with distinctive ERK1/2 activation profiles and gene transcription regulation. In addition, cells expressing IR-B showed higher AKT phosphorylation after insulin stimulation than cells expressing IR-A. Taken together, these results suggest that IR signaling is dependent on localization; internalized IRs regulate mitogenic activity, whereas metabolic balance signaling occurs at the cell membrane.

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Section: Biotinamido-caproyl Insulin (Bac-ins) and Quantum Dots Bind mentioning

confidence: 99%

Differential endocytosis and signaling dynamics of insulin receptor variants IR-A and IR-B

Giudice

Leskow

Arndt‐Jovin

et al. 2011

Journal of Cell Science

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“…Not like in axons of DRG neurons (Cui et al, 2007), in the neurites of PC12 cells, we did not observe biased retrograde transport of fluorescently-labeled NGF. This result was consistent with that of the intracellular transport of quantum-dot-labeled NGF in PC12 cells (Echarte et al, 2007).…”

Section: Endocytosis Of Cy35-ngf In Pc12 Cellssupporting

confidence: 90%

Facilitated intracellular transport of TrkA by an interaction with nerve growth factor

Nomura

Nagai

Harada

et al. 2011

Developmental Neurobiology

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Intracellular transport of neurotrophin receptors together with neurotrophins is one of the key events of neurotrophin signaling for the growth and the survival of neurons. However, the involvement of neurotrophin signaling in the regulation of intracellular transport of neurotrophin receptors has been remained unclear. We visualized the behavior of TrkA, a receptor of nerve growth factor (NGF), by labeling with GFP in PC12 cells. We found remarkable changes of the behavior of TrkA-GFP upon the application of NGF. Before the application, only ~37% of the fluorescent dots of TrkA showed translocations along neurites of PC12 cells. After the application, number of the dots showing the directional movement increased to ~65%. The averaged velocities of the directional movement of TrkA-GFP dots became higher after the application of NGF. We tested the idea whether NGF binding accelerated the translocations of TrkA by simultaneously observing TrkA-GFP and fluorescently labeled NGF, Cy3.5-NGF. The velocity of TrkA-GFP dots associated with Cy3.5-NGF was remarkably higher than that of TrkA-GFP dots without Cy3.5-NGF. On the basis of these observations, we hypothesize that there is a signaling mechanism within a single vesicle that facilitates the intracellular transport of each vesicle containing the activated TrkA.

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“…These conjugates showed superior photostability, lateral resolution, and sensitivity relative to organic dyes. These applications have inspired the use QDs for monitoring other plasma membrane proteins such as integrins [50,66], tyrosine kinases [67,68], G-protein coupled receptors [69], and membrane lipids associated with apoptosis [70,71]. As well, detailed procedures for receptor labeling and visualization of receptor dynamics with QDs have recently been published [72,73], and new techniques to label plasma membrane proteins using versatile molecular biology methods have been developed [74,75].…”

Section: Imaging and Tracking Of Membrane Receptorsmentioning

confidence: 99%

“…Without the ability to wash away unbound probes, which is a crucial step for intracellular labeling of fixed, permeabilized cells, the need for activateable probes that are 'off' until they reach their intended target is apparent. However QDs have already found a niche for quantitative monitoring of motor protein transport and for tracking the fate of internalized receptors, allowing the study of downstream signaling pathways in real time with high signal-to-noise and high temporal and spatial resolution [58,67,68,85,86].…”

Section: Intracellular Delivery Of Qdsmentioning

confidence: 99%

Bioconjugated quantum dots for in vivo molecular and cellular imaging☆

Smith

Duan

Mohs

et al. 2008

Advanced Drug Delivery Reviews

1,086

760

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Semiconductor quantum dots (QDs) are tiny light-emitting particles on the nanometer scale, and are emerging as a new class of fluorescent labels for biology and medicine. In comparison with organic dyes and fluorescent proteins, they have unique optical and electronic properties, with size-tunable light emission, superior signal brightness, resistance to photobleaching, and broad absorption spectra for simultaneous excitation of multiple fluorescence colors. QDs also provide a versatile nanoscale scaffold for designing multifunctional nanoparticles with both imaging and therapeutic functions. When linked with targeting ligands such as antibodies, peptides or small molecules, QDs can be used to target tumor biomarkers as well as tumor vasculatures with high affinity and specificity. Here we discuss the synthesis and development of state-of-the-art QD probes and their use for molecular and cellular imaging. We also examine key issues for in vivo imaging and therapy, such as nanoparticle biodistribution, pharmacokinetics, and toxicology.

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Quantitative single particle tracking of NGF–receptor complexes: Transport is bidirectional but biased by longer retrograde run lengths

Cited by 47 publications

References 50 publications

Differential endocytosis and signaling dynamics of insulin receptor variants IR-A and IR-B

Differential endocytosis and signaling dynamics of insulin receptor variants IR-A and IR-B

Facilitated intracellular transport of TrkA by an interaction with nerve growth factor

Bioconjugated quantum dots for in vivo molecular and cellular imaging☆

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