Electroanalytical Eavesdropping on Single Cell Communication

Kim, Donghyuk; Koseoglu, Secil; Manning, Benjamin; Meyer, Anne-Louise; Haynes, Christy L.

doi:10.1021/ac200666c

Cited by 31 publications

(50 citation statements)

References 66 publications

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“…During this time, the soluble (non-polyphosphate-associated) serotonin present in the granules is released. [25] In contrast, T 1/2 is a measure of the total time of a serotonin secretion event which can be influenced by the opening of the fusion pore, the size of the expanded pore, dissolution of the matrix-bound serotonin, and diffusion of that serotonin into the extracellular space. When stimulated with ionomycin, cow platelets showed slower release kinetics than either rabbit or mouse platelets (larger T rise and T 1/2 values) (Figure 8).…”

Section: Resultsmentioning

confidence: 99%

Platelet membrane variations and their effects on δ-granule secretion kinetics and aggregation spreading among different species

Gruba

Koseoglu

Meyer

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Background Platelet exocytosis is regulated partially by the granular/cellular membrane lipids and proteins. Some platelets contain a membrane-bound tube, called an open canalicular system (OCS), which assists in granular release events and increases the membrane surface area for greater spreading. The OCS is not found in all species, and variations in membrane composition can cause changes in platelet secretion. Since platelet studies use various animal models, it is important to understand how platelets differ in both their composition and granular release to draw conclusions among various models. Methods The relative phospholipid composition of the platelets with (mouse, rabbit) and without (cow) an OCS was quantified using UPLC-MS/MS. Cholesterol and protein composition was measured using an Amplex Red Assay and BCA Assay. TEM and dark field imaging were used to gather platelet images and analyzed with Image J. Granular release was monitored with single cell carbon fiber microelectrode amperometry. Results Cow platelets contained greater amounts of cholesterol and sphingomyelin. In addition, they yield greater serotonin release and longer δ granule secretion times. Finally, they showed greater spreading area with a greater range of spread. Conclusion Platelets containing an OCS had more similarities in their membrane composition and secretion kinetics compared to cow platelets. However, cow platelets showed greater fusion pore stability which could be due to extra sphingomyelin and cholesterol, the primary components of lipid rafts. In addition, their greater stability may lead to more granules assisting in spreading. General Significance Highlights fundamental membrane differences and their effects on platelet secretion

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Section: Resultsmentioning

confidence: 99%

Platelet membrane variations and their effects on δ-granule secretion kinetics and aggregation spreading among different species

Gruba

Koseoglu

Meyer

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…Carbon-fiber microelectrodes have high sensitivity and a low, stable background that facilitates single-cell analysis of MC degranulation. [22–27] During CFMA measurements, a microelectrode is set to an oxidizing potential and placed in contact with a single MC. A micropipette provides local delivery of a stimulating substance used to induce MC degranulation.…”

Section: Introductionmentioning

confidence: 99%

Single-cell analysis of mast cell degranulation induced by airway smooth muscle-secreted chemokines

Manning

Meyer

Gruba

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

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Background Asthma is a chronic inflammatory disease characterized by narrowed airways, bronchial hyper-responsiveness, mucus hyper-secretion, and airway remodeling. Mast cell (MC) infiltration into airway smooth muscle (ASM) is a defining feature of asthma, and ASM regulates the inflammatory response by secreting chemokines, including CXCL10 and CCL5. Single cell analysis offers a unique approach to study specific cellular signaling interactions within large and complex signaling networks such as the inflammatory microenvironment in asthma. Methods Carbon fiber microelectrode amperometry was used to study the effects of ASM–secreted chemokines on mouse peritoneal MC degranulation. Results MC degranulation in response to CXCL10 and CCL5 was monitored at the single cell level. Relative to IgE-mediated degranulation, CXCL10- and CCL5-stimulated MCs released a decreased amount of serotonin per granule with fewer release events per cell. Decreased serotonin released per granule was correlated with increased spike half-width and rise-time values. Conclusions MCs are directly activated with ASM-associated chemokines. CXCL10 and CCL5 induce less robust MC degranulation compared to IgE- and A23187-stimulation. The kinetics of MC degranulation are signaling pathway-dependent, suggesting a biophysical mechanism of regulated degranulation that incorporates control over granule trafficking, transport, and docking machinery. General Significance The biophysical mechanisms, including variations in number of exocytotic release events, serotonin released per granule, and the membrane kinetics of exocytosis that underlie MC degranulation in response to CXCL10 and CCL5 were characterized at the single cell level. These findings clarify the function of ASM-derived chemokines as instigators of MC degranulation relative to classical mechanisms of MC stimulation.

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“…Recently, fluorescent labeling methods have provided many opportunities to study neural networks. However, these methods are not capable of directly detecting and visualizing the neurotransmitter itself but rather utilize the structural changes of synaptic cell membranes and vesicles that contain the neurotransmitters (Kim et al, 2011;Rodriguez et al, 2013). Therefore, direct detection of neurotransmitters with high spatial and temporal resolution is still highly needed.…”

Section: Neurotransmitter Recognition Using Neurosensorsmentioning

confidence: 99%

Nanoparticle‐Templated Molecular Recognition Platforms for Detection of Biological Analytes

Beyene

Demirer

Landry

2016

CP Chemical Biology

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3 These authors contributed equally to this work.Molecular recognition of biological analytes with optical nanosensors provides both spatial and temporal biochemical information. A recently developed sensing platform exploits near-infrared fluorescent single-wall carbon nanotubes combined with electrostatically pinned heteropolymers to yield a synthetic molecular recognition technique that is maximally transparent through biological matter. This molecular recognition technique is known as corona phase molecular recognition (CoPhMoRe). In CoPhMoRe, the specificity of a folded polymer toward an analyte does not arise from a pre-existing polymeranalyte chemical affinity. Rather, specificity is conferred through conformational changes undergone by a polymer that is pinned to the surface of a nanoparticle in the presence of an analyte and the subsequent modifications in fluorescence readout of the nanoparticles. The protocols in this article describe a novel single-molecule microscopy tool (near-infrared fluorescence and total internal reflection fluorescence [nIRF TIRF] hybrid microscope) to visualize the CoPhMoRe recognition process, enabling a better understanding of synthetic molecular recognition. We describe this requisite microscope for simultaneous single-molecule visualization of optical molecular recognition and signal transduction. We elaborate on the general procedures for synthesizing and identifying single-walled carbon nanotube-based sensors that employ CoPhMoRe via two biologically relevant examples of single-molecule recognition for the hormone estradiol and the neurotransmitter dopamine. C 2016 by John Wiley & Sons, Inc.Keywords: fluorescence microscopy r molecular recognition r near-infrared imaging r nanoparticles r neurotransmitter r nIRF TIRF hybrid microscope r single-walled carbon nanotube (SWCNT) r screening r single molecule imaging r sensors How to cite this article: Beyene, A.G., Demirer, G.S., and Landry, M.P. 2016. Nanoparticle-templated molecular recognition platforms for detection of biological analytes Curr.

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Electroanalytical Eavesdropping on Single Cell Communication

Cited by 31 publications

References 66 publications

Platelet membrane variations and their effects on δ-granule secretion kinetics and aggregation spreading among different species

Platelet membrane variations and their effects on δ-granule secretion kinetics and aggregation spreading among different species

Single-cell analysis of mast cell degranulation induced by airway smooth muscle-secreted chemokines

Nanoparticle‐Templated Molecular Recognition Platforms for Detection of Biological Analytes

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