A multiscale study of the role of dynamin in the regulation of glucose uptake

Trouillon, Raphaël; Letizia, Maria Cristina; Menzies, Keir J.; Mouchiroud, Laurent; Auwerx, Johan; Gijs, Martin A. M.

doi:10.1039/c7ib00015d

Cited by 8 publications

(10 citation statements)

References 64 publications

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“…Additionally, the deleterious effects of NaCl are greater than those of sucrose at the same molarity 5 . Additional phenotypic results also revealed that high doses of glucose severely affect the motility of nematodes 14 .…”

Section: Introductionmentioning

confidence: 94%

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Automated phenotyping of Caenorhabditis elegans embryos with a high-throughput-screening microfluidic platform

et al. 2020

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The nematode Caenorhabditis elegans has been extensively used as a model multicellular organism to study the influence of osmotic stress conditions and the toxicity of chemical compounds on developmental and motilityassociated phenotypes. However, the several-day culture of nematodes needed for such studies has caused researchers to explore alternatives. In particular, C. elegans embryos, due to their shorter developmental time and immobile nature, could be exploited for this purpose, although usually their harvesting and handling is tedious. Here, we present a multiplexed, high-throughput and automated embryo phenotyping microfluidic approach to observe C. elegans embryogenesis after the application of different chemical compounds. After performing experiments with up to 800 embryos per chip and up to 12 h of time-lapsed imaging per embryo, the individual phenotypic developmental data were collected and analyzed through machine learning and image processing approaches. Our proof-of-concept platform indicates developmental lag and the induction of mitochondrial stress in embryos exposed to high doses (200 mM) of glucose and NaCl, while small doses of sucrose and glucose were shown to accelerate development. Overall, our new technique has potential for use in large-scale developmental biology studies and opens new avenues for very rapid high-throughput and high-content screening using C. elegans embryos.

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Section: Introductionmentioning

confidence: 94%

“…L-Glucose is not biologically active, and this compound affects the nematode differently. Previous studies determined the different impacts of these two enantiomer compounds on lifespan 39 , motility 14 , the egg laying rate 40 , and nematode sensitivity 41 .…”

Section: Osmotic Influence On Embryo Developmentmentioning

confidence: 99%

Automated phenotyping of Caenorhabditis elegans embryos with a high-throughput-screening microfluidic platform

et al. 2020

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“…2-DG transport -into Soleus and EDL muscles were assessed as described before (80). Electrochemical glucose sensing -Inspired by (81) we fabricated a microfluidic chip system using standard soft lithographic techniques. The chips were divided into a tissue chamber and a glucose sensing chamber connected by tubing.…”

Section: Glucose Uptake Measurementsmentioning

confidence: 99%

“…Next the fiber-containing paper patch were inserted into the tissue chamber and perifused with serum free DMEM containing 4 mM glucose for 3-5 minutes. Next, the glucose concentration in the perfusate was monitored during basal and insulin-stimulated conditions and ∆glucose was calculated as previously described (81). Colchicine and Noco treatment for 2h was achieved by preincubating the paper patches containing fibers in colchicine and Noco and keeping the drugs in the perfusate at all times after the assembly of the tissue chamber.…”

Section: Glucose Uptake Measurementsmentioning

confidence: 99%

Microtubule-mediated GLUT4 trafficking is disrupted in insulin resistant skeletal muscle

Knudsen

Persson

Henríquez‐Olguín

et al. 2022

Preprint

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Microtubules serve as tracks for long-range intracellular trafficking of glucose transporter 4 (GLUT4), but the role of this process in skeletal muscle and insulin resistance is unclear. Here, we used fixed and live-cell imaging to study microtubule-based GLUT4 trafficking in human and mouse muscle fibers and L6 rat muscle cells. We found GLUT4 localized along and on the microtubules in mouse and human muscle fibers. Pharmacological microtubule disruption using Nocodazole (Noco) prevented long-range GLUT4 trafficking and depleted GLUT4-enriched structures at microtubule nucleation sites in a fully reversible manner. Using a perfused muscle-on-a-chip system to enable real-time glucose uptake measurements in isolated mouse skeletal muscle fibers, we observed that Noco maximally disrupted the microtubule network after 5 min without affecting insulin-stimulated glucose uptake. In contrast, a 2h Noco treatment markedly decreased insulin responsiveness of glucose uptake. Insulin resistance in mouse muscle fibers induced either in vitro by C2 ceramides or in vivo by diet-induced obesity, impaired microtubule-based GLUT4 trafficking. In L6 muscle cells, pharmacological activation of the microtubule motor protein kinesin-1 increased basal and insulin-stimulated GLUT4 translocation, whereas shRNA-mediated knockdown of the kinesin-1 protein encoding gene Kif5B reduced insulin-stimulated GLUT4 translocation. Thus, in adult skeletal muscle fibers, the microtubule network is essential for intramyocellular GLUT4 movement, likely functioning to maintain an insulin-responsive cell-surface recruitable GLUT4 pool via kinesin-1 mediated trafficking.

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“…Its high porosity facilitates wicking and makes it highly amenable to microfluidic applications and fluid handling . Its chemical inertness allows for the culture of cells, thus providing a robust substrate for three‐dimensional multicellular assays …”

Section: Introductionmentioning

confidence: 99%

Paper‐Based Polymer Electrodes for Bioanalysis and Electrochemistry of Neurotransmitters

Trouillon

Gijs

2018

ChemPhysChem

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This article describes the preparation and characterization of PEDOT-coated paper electrodes. Their specific behavior was investigated, especially the impact of electrode shape on the electrochemical properties. It was found that different electrode geometries promote different results, largely because of a potential drop in the bulk of the electrode material. More importantly, the suitability of these substrates for bio- and neurochemical analyses was investigated. The paper electrodes were found to offer better resistance to both protein and neurotransmitter foulings, in comparison to a planar electrode. Interestingly, long paper electrodes were more stable during sustained oxidation of dopamine and serotonin than short ones, possibly because of the conjunction of surface passivation and potential drop allowing for the site of oxidation to move along the electrode as fouling progresses.

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A multiscale study of the role of dynamin in the regulation of glucose uptake

Cited by 8 publications

References 64 publications

Automated phenotyping of Caenorhabditis elegans embryos with a high-throughput-screening microfluidic platform

Automated phenotyping of Caenorhabditis elegans embryos with a high-throughput-screening microfluidic platform

Microtubule-mediated GLUT4 trafficking is disrupted in insulin resistant skeletal muscle

Paper‐Based Polymer Electrodes for Bioanalysis and Electrochemistry of Neurotransmitters

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