Super-Resolution Imaging and Spatial Analysis of RAS on Intact Plasma Membrane Sheets

2021 ◽  
pp. 217-232
Author(s):  
Yong Zhou ◽  
John F. Hancock
Keyword(s):  
Plasma Membrane ◽  
Spatial Analysis ◽  
Super Resolution ◽  
Resolution Imaging

scholarly journals Super-Resolution Imaging of Plasma Membrane Lesions Inflicted by 405-nm Laser Light

2016 ◽  
Vol 110 (3) ◽  
pp. 488a
Author(s):  
Lu Zhou ◽  
Volker Middel ◽  
G. Ulrich Nienhaus ◽  
Uwe Uwe Strähle
Keyword(s):  
Plasma Membrane ◽  
Laser Light ◽  
Super Resolution ◽  
Resolution Imaging

scholarly journals Nanobodies reveal an extra-synaptic population of SNAP-25 and Syntaxin 1A in hippocampal neurons

2018 ◽  
Author(s):  
Manuel Maidorn ◽  
Aurélien Olichon ◽  
Silvio O. Rizzoli ◽  
Felipe Opazo
Keyword(s):  
Plasma Membrane ◽  
Synaptic Vesicle ◽  
Hippocampal Neurons ◽  
Cellular Localization ◽  
Super Resolution ◽  
Syntaxin 1A ◽  
Resolution Imaging ◽  
Single Domain Antibodies ◽  
The Subject ◽  
Synaptic Vesicle Fusion

AbstractSynaptic vesicle fusion (exocytosis) is a precisely regulated process that entails the formation of SNARE complexes between the vesicle protein synaptobrevin 2 (VAMP2) and the plasma membrane proteins Syntaxin 1 and SNAP-25. The sub-cellular localization of the latter two molecules remains unclear, although they have been the subject of many recent investigations. To address this, we generated two novel camelid single domain antibodies (nanobodies) specifically binding to SNAP-25 and Syntaxin 1A. These probes penetrated more easily into samples and detected their targets more efficiently than conventional antibodies in crowded regions. When investigated by super-resolution imaging, the nanobodies revealed substantial extra-synaptic populations for both SNAP-25 and Syntaxin 1A, which were poorly detected by antibodies. Moreover, extra-synaptic Syntaxin 1A molecules were recruited to synapses during stimulation, suggesting that these are physiologically-active molecules. We conclude that nanobodies are able to reveal qualitatively and quantitatively different organization patterns, when compared to conventional antibodies.

scholarly journals Microtubules Regulate Spatial Localization and Availability of Insulin Granules in Pancreatic Beta Cells

2019 ◽  
Author(s):  
Kai M. Bracey ◽  
Kung-Hsien Ho ◽  
Dmitry Yampolsky ◽  
Guoqiang Gu ◽  
Irina Kaverina ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Beta Cells ◽  
Pancreatic Beta Cells ◽  
Single Cells ◽  
Critical Role ◽  
Super Resolution ◽  
Transport Dynamics ◽  
Insulin Granules ◽  
Resolution Imaging ◽  
Glucose Stimulated Insulin Secretion

AbstractTwo key prerequisites for glucose stimulated insulin secretion (GSIS) in Beta cells are the proximity of insulin granules to the plasma membrane and their anchoring or docking to the plasma membrane (PM). While recent evidence has indicated that both of these factors are altered in the context of diabetes, it is unclear what regulates localization of insulin and its interactions with the PM within single cells. Here we demonstrate that microtubule (MT) motor mediated transport dynamics have a critical role in regulating both factors. Super-resolution imaging shows that while the MT cytoskeleton resembles a random meshwork in the cells’ interior, MTs near the cells surface are preferentially aligned with the PM. Computational modeling demonstrates two consequences of this alignment. First, this structured MT network preferentially withdraws granules from the PM. Second, the binding and transport of insulin granules by MT motors prevents their stable anchoring to the PM. The MT cytoskeleton thus negatively regulates GSIS by both limiting the amount of insulin proximal to the PM and preventing/breaking interactions between the PM and the remaining nearby insulin. These results predict that altering MT structure in beta cells can be used to tune GSIS. Thus, our study points to a potential of an alternative therapeutic strategy for diabetes by targeting specific MT regulators.

Super-Resolution Imaging of Plasma Membrane Glycans

2014 ◽  
Vol 53 (41) ◽  
pp. 10921-10924 ◽  
Author(s):  
Sebastian Letschert ◽  
Antonia Göhler ◽  
Christian Franke ◽  
Nadja Bertleff-Zieschang ◽  
Elisabeth Memmel ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Super Resolution ◽  
Resolution Imaging

Super-Resolution Imaging of Plasma Membrane Glycans

2014 ◽  
Vol 126 (41) ◽  
pp. 11101-11104 ◽  
Author(s):  
Sebastian Letschert ◽  
Antonia Göhler ◽  
Christian Franke ◽  
Nadja Bertleff-Zieschang ◽  
Elisabeth Memmel ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Super Resolution ◽  
Resolution Imaging

scholarly journals Local palmitoylation cycles define activity-regulated postsynaptic subdomains

2013 ◽  
Vol 202 (1) ◽  
pp. 145-161 ◽  
Author(s):  
Yuko Fukata ◽  
Ariane Dimitrov ◽  
Gaelle Boncompain ◽  
Ole Vielemeyer ◽  
Franck Perez ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Dendritic Spines ◽  
Recombinant Antibody ◽  
Super Resolution ◽  
Live Cell ◽  
Synaptic Activity ◽  
Membrane Targeting ◽  
Resolution Imaging

Distinct PSD-95 clusters are primary landmarks of postsynaptic densities (PSDs), which are specialized membrane regions for synapses. However, the mechanism that defines the locations of PSD-95 clusters and whether or how they are reorganized inside individual dendritic spines remains controversial. Because palmitoylation regulates PSD-95 membrane targeting, we combined a conformation-specific recombinant antibody against palmitoylated PSD-95 with live-cell super-resolution imaging and discovered subsynaptic nanodomains composed of palmitoylated PSD-95 that serve as elementary units of the PSD. PSD-95 in nanodomains underwent continuous de/repalmitoylation cycles driven by local palmitoylating activity, ensuring the maintenance of compartmentalized PSD-95 clusters within individual spines. Plasma membrane targeting of DHHC2 palmitoyltransferase rapidly recruited PSD-95 to the plasma membrane and proved essential for postsynaptic nanodomain formation. Furthermore, changes in synaptic activity rapidly reorganized PSD-95 nano-architecture through plasma membrane–inserted DHHC2. Thus, the first genetically encoded antibody sensitive to palmitoylation reveals an instructive role of local palmitoylation machinery in creating activity-responsive PSD-95 nanodomains, contributing to the PSD (re)organization.

Triple-Color Super-Resolution Imaging of Live Cells: Resolving Submicroscopic Receptor Organization in the Plasma Membrane

2012 ◽  
Vol 51 (20) ◽  
pp. 4868-4871 ◽  
Author(s):  
Stephan Wilmes ◽  
Markus Staufenbiel ◽  
Domenik Liße ◽  
Christian P. Richter ◽  
Oliver Beutel ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Super Resolution ◽  
Live Cells ◽  
Resolution Imaging

Triple-Color Super-Resolution Imaging of Live Cells: Resolving Submicroscopic Receptor Organization in the Plasma Membrane

2012 ◽  
Vol 124 (20) ◽  
pp. 4952-4955 ◽  
Author(s):  
Stephan Wilmes ◽  
Markus Staufenbiel ◽  
Domenik Liße ◽  
Christian P. Richter ◽  
Oliver Beutel ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Super Resolution ◽  
Live Cells ◽  
Resolution Imaging
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