Single-Molecule Fluorescence Measurement of SNARE-Mediated Vesicle Fusion

Hu, Yachong; Tian, Ze; Diao, Jiajie

doi:10.1007/978-1-4939-8760-3_22

Cited by 5 publications

(6 citation statements)

References 21 publications

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“…The sequence of fusion intermediates from lipid monolayers to a complete bilayer merge has been reported in vacuoles 192 . Hu et al correlated the membrane fusion stages with a molecular mechanism using reconstituted vesicles 193 . At a single-vesicle level, they traced the lipid-mixing process using FRET microscopy and correlated it with the docking, hemifusion and full-fusion stages 193 .…”

Section: Ev Characterizationmentioning

confidence: 99%

“…Hu et al correlated the membrane fusion stages with a molecular mechanism using reconstituted vesicles 193 . At a single-vesicle level, they traced the lipid-mixing process using FRET microscopy and correlated it with the docking, hemifusion and full-fusion stages 193 . They also report that an optimal distance for a SNARE-mediated fusion is 5 nm.…”

Section: Ev Characterizationmentioning

confidence: 99%

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Using single-vesicle technologies to unravel the heterogeneity of extracellular vesicles

et al. 2021

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Extracellular vesicles (EVs) are heterogeneous lipid containers with a complex molecular cargo comprising several populations with unique roles in biological processes. These vesicles are closely associated with specific physiological features, which makes them invaluable in the detection and monitoring of various diseases. EVs play a key role in pathophysiological processes by actively triggering genetic or metabolic responses. However, the heterogeneity of their structure and composition hinders their application in medical diagnosis and therapies. This diversity makes it difficult to establish their exact physiological roles, and the functions and composition of different EV (sub)populations. Ensemble averaging approaches currently employed for EV characterization, such as western blotting or 'omics' technologies, tend to obscure rather than reveal these heterogeneities. Recent developments in single-vesicle analysis have made it possible to overcome these limitations and have facilitated the development of practical clinical applications. In this review, we discuss the benefits and challenges inherent to the current methods for the analysis of single vesicles and review the contribution of these approaches to the understanding of EV biology. We describe the contributions of these recent technological advances to the characterization and phenotyping of EVs, examination of the role of EVs in cell-to-cell communication pathways and the identification and validation of EVs as disease biomarkers. Finally, we discuss the potential of innovative single-vesicle imaging and analysis methodologies using microfluidic devices, which promise to deliver rapid and effective basic and practical applications for minimally invasive prognosis systems.

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Section: Ev Characterizationmentioning

confidence: 99%

Section: Ev Characterizationmentioning

confidence: 99%

Using single-vesicle technologies to unravel the heterogeneity of extracellular vesicles

et al. 2021

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“…With the further understanding of the synaptic vesicle cycle, single-molecule smFRET imaging technology will be needed to monitor the interaction between proteins in real time to explore SNAREmediated membrane fusion (118,(135)(136)(137). It is expected that the precise role of related proteins in neurotransmitter release will be further expanded with the application of advanced bioanalytical techniques.…”

Section: Discussionmentioning

confidence: 99%

Research progress on vesicle cycle and neurological disorders

2021

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Neurons are special polarized cells whose synaptic vesicles release neurotransmitters into the synaptic cleft, acting on postsynaptic receptors and thus transmitting information from presynaptic to postsynaptic states. The integrity of the vesicle cycle is critical to the transmission of neural signals in the brain. According to the molecular mechanism of calcium-triggered release, the assembly of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) is required in the process of synaptic vesicle fusion and vesicle exocytosis. Many delicate steps are required to maintain the dynamic process of ‘release-recycle’, including intermediate processes and the dynamic balance of neurotransmission. Various neurodegenerative and neuropsychiatric diseases result from synaptic cycle dysfunction. This review of the relationships between the structure and function of synaptic vesicles in physiological and pathological conditions provides a theoretical basis for synaptic transmission and a novel avenue for the study of synaptic plasticity associated with mood disorders, highlighting potential targets for treating diseases.

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“…DiO, for instance, absorbs strongly at 484 nm with peak emission at 501 nm, while DiI and DiD absorb and emit at 549/644 nm and 565/665 nm, respectively (Figure 3b). Much like the short-chain derivatives, the longer chains also have high extinction coefficients (DiO ~ 140,000 M -1 cm -1 ; Dil ~ 148,000 M -1 cm -1 ; DiD ~ 193,000) and comparable excited state lifetimes (~ 1 ns), though their quantum yields tend to be somewhat lower (~0.07) (57,58) . Such probes offer a chance to probe the local dynamics and curvature of lipid bilayers via measurement of their rotational time trajectories (42,59) .…”

Section: Organic Dyesmentioning

confidence: 99%

A guide to small fluorescent probes for single-molecule biophysics

Leake

Quinn

2023

Chemical Physics Reviews

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The explosive growth of single-molecule techniques is transforming our understanding of biology, helping to develop new physics inspired by emergent biological processes, and leading to emerging areas of nanotechnology. Key biological and chemical processes can now be probed with new levels of detail, one molecule at a time, from the nanoscopic dynamics of nature's molecular machines to an ever-expanding range of exciting applications across multiple length and time scales. Their common feature is an ability to render the underlying distribution of molecular properties that ensemble averaging masks and to reveal new insights into complex systems containing spatial and temporal heterogeneity. Small fluorescent probes are among the most adaptable and versatile for single-molecule sensing applications because they provide high signal-to-noise ratios combined with excellent specificity of labeling when chemically attached to target biomolecules or embedded within a host material. In this review, we examine recent advances in probe designs, their utility, and applications and provide a practical guide to their use, focusing on the single-molecule detection of nucleic acids, proteins, carbohydrates, and membrane dynamics. We also present key challenges that must be overcome to perform successful single-molecule experiments, including probe conjugation strategies, identify tradeoffs and limitations for each probe design, showcase emerging applications, and discuss exciting future directions for the community.

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Single-Molecule Fluorescence Measurement of SNARE-Mediated Vesicle Fusion

Cited by 5 publications

References 21 publications

Using single-vesicle technologies to unravel the heterogeneity of extracellular vesicles

Using single-vesicle technologies to unravel the heterogeneity of extracellular vesicles

Research progress on vesicle cycle and neurological disorders

A guide to small fluorescent probes for single-molecule biophysics

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