Probing labeling-induced lysosome alterations in living cells by imaging-derived mean squared displacement analysis

Durso, William; D’Autilia, Francesca; Amodeo, Rosy; Marchetti, Laura; Cardarelli, Francesco

doi:10.1016/j.bbrc.2018.08.028

Cited by 4 publications

(4 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…11). Worthy of mention, present results on granule enlargement are corroborated by analogous observations made by us on a different sub-cellular organelle, the lysosome, under over-expression of GFP-tagged transmembrane protein markers 36 . The effect on granule dynamic properties, on the other hand, prompted us to speculate on a possible effect of phogrin-FP overload on the correct granule docking to the trafficking machinery (e.g.…”

Section: Resultssupporting

confidence: 88%

Insulin secretory granules labelled with phogrin-fluorescent proteins show alterations in size, mobility and responsiveness to glucose stimulation in living β-cells

Ferri

Digiacomo

Lavagnino

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

The intracellular life of insulin secretory granules (ISGs) from biogenesis to secretion depends on their structural (e.g. size) and dynamic (e.g. diffusivity, mode of motion) properties. Thus, it would be useful to have rapid and robust measurements of such parameters in living β-cells. To provide such measurements, we have developed a fast spatiotemporal fluctuation spectroscopy. We calculate an imaging-derived Mean Squared Displacement ( i MSD), which simultaneously provides the size, average diffusivity, and anomalous coefficient of ISGs, without the need to extract individual trajectories. Clustering of structural and dynamic quantities in a multidimensional parametric space defines the ISGs’ properties for different conditions. First, we create a reference using INS-1E cells expressing proinsulin fused to a fluorescent protein (FP) under basal culture conditions and validate our analysis by testing well-established stimuli, such as glucose intake, cytoskeleton disruption, or cholesterol overload. After, we investigate the effect of FP-tagged ISG protein markers on the structural and dynamic properties of the granule. While i MSD analysis produces similar results for most of the lumenal markers, the transmembrane marker phogrin-FP shows a clearly altered result. Phogrin overexpression induces a substantial granule enlargement and higher mobility, together with a partial de-polymerization of the actin cytoskeleton, and reduced cell responsiveness to glucose stimulation. Our data suggest a more careful interpretation of many previous ISG-based reports in living β-cells. The presented data pave the way to high-throughput cell-based screening of ISG structure and dynamics under various physiological and pathological conditions.

show abstract

Section: Resultssupporting

confidence: 88%

Insulin secretory granules labelled with phogrin-fluorescent proteins show alterations in size, mobility and responsiveness to glucose stimulation in living β-cells

Ferri

Digiacomo

Lavagnino

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…At this point, the characteristic parameters describing ISG average structural properties (i.e., size) and dynamics (i.e., diffusion coefficient, D, and anomalous coefficient, α, at different spatiotemporal scales) can be readily extracted and used to define the fingerprint of the structure of interest. The potential of the method was already demonstrated for a variety of biological objects, ranging from molecules to nanoparticles or entire subcellular organelles/structures [ 22 , 23 , 24 , 25 , 26 , 27 , 28 ]. In particular, some of us recently validated the i MSD approach to the use of fluorescently labeled ISGs in a model of β-like immortalized cells, the insulinoma-1E (INS-1E) cells [ 26 ].…”

Section: Introductionmentioning

confidence: 99%

Spatiotemporal Correlation Spectroscopy Reveals a Protective Effect of Peptide-Based GLP-1 Receptor Agonism against Lipotoxicity on Insulin Granule Dynamics in Primary Human β-Cells

et al. 2021

Self Cite

View full text Add to dashboard Cite

Glucagon-like peptide-1 receptor (GLP-1R) agonists are being used for the treatment of type 2 diabetes (T2D) and may have beneficial effects on the pancreatic β-cells. Here, we evaluated the effects of GLP-1R agonism on insulin secretory granule (ISG) dynamics in primary β-cells isolated from human islets exposed to palmitate-induced lipotoxic stress. Islets cells were exposed for 48 h to 0.5 mM palmitate (hereafter, ‘Palm’) with or without the addition of a GLP-1 agonist, namely 10 nM exendin-4 (hereafter, ‘Ex-4′). Dissociated cells were first transfected with syncollin-EGFP in order to fluorescently mark the ISGs. Then, by applying a recently established spatiotemporal correlation spectroscopy technique, the average structural (i.e., size) and dynamic (i.e., the local diffusivity and mode of motion) properties of ISGs are extracted from a calculated imaging-derived Mean Square Displacement (iMSD) trace. Besides defining the structural/dynamic fingerprint of ISGs in human cells for the first time, iMSD analysis allowed to probe fingerprint variations under selected conditions: namely, it was shown that Palm affects ISGs dynamics in response to acute glucose stimulation by abolishing the ISGs mobilization typically imparted by glucose and, concomitantly, by reducing the extent of ISGs active/directed intracellular movement. By contrast, co-treatment with Ex-4 normalizes ISG dynamics, i.e., re-establish ISG mobilization and ability to perform active transport in response to glucose stimulation. These observations were correlated with standard glucose-stimulated insulin secretion (GSIS), which resulted in being reduced in cells exposed to Palm but preserved in cells concomitantly exposed to 10 nM Ex-4. Our data support the idea that GLP-1R agonism may exert its beneficial effect on human β-cells under metabolic stress by maintaining ISGs’ proper intracellular dynamics.

show abstract

“…The MSD curve of dye molecules can be achieved by MD to simulate the movement path of dye molecules in the process of movement. 41,42 The MSD curve relationship can be expressed as MSD=〈|r(t)−r(0)|2〉…”

Section: Methodsmentioning

confidence: 99%

Level dyeing property and aggregation morphology of reactive black 5 on cotton fabric in a salt-free and less-water dyeing system

Hu,

Pei,

Shen

et al. 2023

Textile Research Journal

View full text Add to dashboard Cite

In a traditional water-based dyeing system, large amounts of salts and water are consumed. However, a non-aqueous media dyeing system achieves salt-free and less-water dyeing. In a non-aqueous media dyeing system, a small amount of water is used, which may affect the aggregation morphology of the dyes. The molecular dynamics of reactive dyes on the surface of cotton fibers were investigated. The results showed that the color depth of dyed cotton fabric did not change much when the dosage of salt was below 5.2% o.w.f. However, continuing to increase the amount of salt could influence the dyeing level of dyed fabric. The diffusion coefficient of dye was 0.3044 × 10−5 cm−2/s when there was no salt in dyeing system. But it was decreased by 14.03% and 16.62% when the salt concentration was increased from zero to 5.2% and 13% o.w.f., respectively. When the concentration of sodium sulfate was zero, the dye molecules in the solution displayed in the form of unimolecular molecules and dimers. However, reactive dyes mainly existed in the form of trimers and polymers when the concentration of sodium sulfate was above 5.2%. When there was no salt in the less-water dyeing system, the van der Waal energy between fiber and dye was 131.29 kJ/mol, but it was increased to 382.92 kJ/mol when the dosage of sodium sulfate was 5.2%. Therefore, with the increase of sodium sulfate dosage, the color depth of the dyed fabric changed little, but the level of dyeing property became poorer.

show abstract

Probing labeling-induced lysosome alterations in living cells by imaging-derived mean squared displacement analysis

Cited by 4 publications

References 22 publications

Insulin secretory granules labelled with phogrin-fluorescent proteins show alterations in size, mobility and responsiveness to glucose stimulation in living β-cells

Insulin secretory granules labelled with phogrin-fluorescent proteins show alterations in size, mobility and responsiveness to glucose stimulation in living β-cells

Spatiotemporal Correlation Spectroscopy Reveals a Protective Effect of Peptide-Based GLP-1 Receptor Agonism against Lipotoxicity on Insulin Granule Dynamics in Primary Human β-Cells

Level dyeing property and aggregation morphology of reactive black 5 on cotton fabric in a salt-free and less-water dyeing system

Contact Info

Product

Resources

About