A5065272998- Lipid Membrane Structure and Behavior
- Advanced Fluorescence Microscopy Techniques
- Receptor Mechanisms and Signaling
- Monoclonal and Polyclonal Antibodies Research
- Cellular transport and secretion
- Force Microscopy Techniques and Applications
- Neuropeptides and Animal Physiology
- Advanced Electron Microscopy Techniques and Applications
- Cell Adhesion Molecules Research
- Neuroscience and Neuropharmacology Research
- Nanopore and Nanochannel Transport Studies
- Cellular Mechanics and Interactions
- RNA Research and Splicing
- Molecular Sensors and Ion Detection
- RNA Interference and Gene Delivery
- Click Chemistry and Applications
- Biotin and Related Studies
- Advanced Biosensing Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Pharmacological Receptor Mechanisms and Effects
- RNA and protein synthesis mechanisms
- Mast cells and histamine
- Digital Holography and Microscopy
- Microfluidic and Bio-sensing Technologies
- Electrohydrodynamics and Fluid Dynamics
Okinawa Institute of Science and Technology Graduate University
2017-2024
Kyoto University
2011-2019
Material Sciences (United States)
2011-2014
Niigata University
2009
The mechanisms by which the diffusion rate in plasma membrane (PM) is regulated remain unresolved, despite their importance spatially regulating reaction rates PM. Proposed models include entrapment nanoscale noncontiguous domains found PtK2 cells, slow due to crowding, and actin-induced compartmentalization. Here, applying single-particle tracking at high time resolutions, mainly PtK2-cell PM, we confined plus hop movements (termed "hop diffusion") for both a nonraft phospholipid...
Neuronal transmission relies on the regulated secretion of neurotransmitters, which are packed in synaptic vesicles (SVs). Hundreds SVs accumulate at boutons. Despite being held together, highly mobile, so that they can be recruited to plasma membrane for their rapid release during neuronal activity. However, how such confinement corroborates with motility remains unclear. To bridge this gap, we employ ultrafast single-molecule tracking (SMT) reconstituted system native and living neurons....
The spatial resolution of fluorescence microscopy has recently been greatly enhanced. However, improvements in temporal have limited, despite their importance for examining living cells. Here, we developed an ultrafast camera system that enables the highest time resolutions single fluorescent-molecule imaging to date, which were photon-limited by fluorophore photophysics: 33 and 100 µs with single-molecule localization precisions 34 20 nm, respectively, Cy3, optimal identified. Using...
Using our newly developed ultrafast camera described in the companion paper, we reduced data acquisition periods required for photoactivation/photoconversion localization microscopy (PALM, using mEos3.2) and direct stochastic reconstruction (dSTORM, HMSiR) by a factor of ≈30 compared with standard methods, much greater view-fields, precisions 29 19 nm, respectively, thus opening up previously inaccessible spatiotemporal scales to cell biology research. Simultaneous two-color PALM-dSTORM...
Using single-molecule imaging with enhanced time resolutions down to 5 ms, we found that CD59 cluster rafts and GM1 were stably induced in the outer leaflet of plasma membrane (PM), which triggered activation Lyn, H-Ras, ERK continually recruited Lyn H-Ras right beneath them inner dwell lifetimes <0.1 s. The detection was possible due employed here. recruitment depended on PM cholesterol saturated alkyl chains whereas it blocked by nonraftophilic transmembrane protein moiety...
Two very polarized views exist for understanding the cellular plasma membrane (PM). For some, it is simple fluid described by original Singer-Nicolson mosaic model. others, due to presence of thousands molecular species that extensively interact with each other, PM forms various clusters and domains are constantly changing therefore, no rules can explain structure dynamics PM. In this article, we propose viewing from its two predominant components, cholesterol actin filaments, provides an...
Abstract The number and subunit compositions of AMPA receptors (AMPARs), hetero- or homotetramers composed four subunits GluA1–4, in the synapse is carefully tuned to sustain basic synaptic activity. This enables stimulation-induced plasticity, which central learning memory. AMPAR tetramers have been widely believed be stable from their formation endoplasmic reticulum until proteolytic decomposition. However, by observing GluA1 GluA2 at level single molecules, we find that homo-...
A key issue in neuronal circuit regulation is how synapse formation initiated. Synapse could start when one or more synaptic scaffold proteins that can initiate reach certain threshold concentrations the dendritic shaft, which might lead to their oligomerization even liquid-liquid phase separation (LLPS). By combining vitro reconstitution of purified with live-cell single-molecule and confocal imaging, we demonstrated SynGAP alone forms assemblies nanoscale clusters containing several tens...
Electron tomography of the plasma membrane (PM) identified several layers cortical actin meshwork running parallel to PM cytoplasmic surface throughout PM. Here, structures and dynamics were examined in living cells, using super-resolution microscopy, with (x,y)- z-resolutions ~140 ~400 nm, respectively, single-molecule imaging. The microscopy sub-micron-sized clusters that appeared identical by both phalloidin post-fixation staining Lifeact-mGFP expression followed fixation, therefore,...
Opioid receptors (ORs) are critical for endogenous and synthetic analgesics. Their homodimerization is considered important their pharmacological diversities, but whether they form homodimers remains controversial. Here, we established that the three classical ORs, mu-, kappa-, delta-ORs (MOR, KOR, DOR, respectively) undergo repeated transient (120-180 ms) homodimerizations every few seconds. This was done by using single-molecule imaging developing theories analyzing colocalization data,...
Abstract Heterodimerization of opioid receptors (ORs), MOR, KOR, and DOR, is implied in their functional regulation diversification, thus its understanding crucial for developing better analgesic treatments. However, our knowledge on OR heterodimerization/heterodimers remains limited. Here, using single-molecule imaging analysis, we found that the main morphine receptor, repeatedly forms transient (≈250 ms) heterodimers with DOR every 1-10 seconds, but not whereas KOR also form heterodimers....
The central mechanism for the transmission of prion protein misfolding is structural conversion normal cellular to pathogenic misfolded protein, by interaction with protein. This process might be enhanced due homo-dimerization/oligomerization However, behaviors in plasma membrane have remained largely unknown. Here, using single fluorescent-molecule imaging, we found that both and Thy1, a control glycosylphosphatidylinositol-anchored exhibited very similar intermittent transient...
SUMMARY Crosstalk of cellular signaling pathways is essential for coordinating cell responses. Therefore, we suspect the existence special scaffolding platforms that specifically undertake signal crosstalk and integration. Using advanced single-molecule imaging, found a nanometer-scale liquid-like platform integrating signals downstream from GPI-anchored receptors receptor-type tyrosine kinases. The employs some focal adhesion proteins, including integrin, talin, RIAM, VASP, zyxin, but...
Abstract Using the ultrafast camera system and new theories for hop diffusion described in companion paper, we first time demonstrated that membrane molecules undergo among compartments bulk basal plasma (PM), with virtually same compartment sizes (108 nm) as those apical PM dwell lifetimes within a (10 24 ms phospholipid transferrin receptor, respectively), suggesting basic structures molecular dynamics are very similar regions of PMs. Ultrafast PALM single-molecule imaging revealed focal...
The immune system has been studied extensively by increasing the demand for object tracking of particles. However, because researches on single-particle (SPT) machine learning have not progressed yet, currently there is a reliance software analysis despite low accuracy. There are three problems with SPT. First, no differences in feature each molecule, so possible. Second, it difficult to predict direction molecular motion random. Third, high density molecules causes frequent ID switches....
<title>Abstract</title> Opioid receptors (ORs) are critical for endogenous and synthetic analgesics. Their homodimerization is considered important their pharmacological diversities, but whether they form homodimers remains controversial. Here, we established that the three classical ORs, mu-, kappa-, delta-ORs (MOR, KOR, DOR, respectively) undergo repeated transient (120-180 ms) homodimerizations every few seconds. This was done by using single-molecule imaging developing theories analyzing...
<title>Abstract</title> Heterodimerization of opioid receptors (ORs), MOR, KOR, and DOR, is implied in their functional regulation diversification, thus its understanding crucial for developing better analgesic treatments. However, our knowledge on OR heterodimerization/heterodimers remains limited. Here, using single-molecule imaging analysis, we found that the main morphine receptor, repeatedly forms <italic>transient (≈250 ms) heterodimers</italic> with DOR every 1-10 seconds, but not...