scholarly journals Live Cell Imaging and 3D Analysis of Angiotensin Receptor Type 1a Trafficking in Transfected Human Embryonic Kidney Cells Using Confocal Microscopy

10.3791/55177 ◽  
2017 ◽  
Author(s):  
Parnika Kadam ◽  
Ryan McAllister ◽  
Jeffrey S. Urbach ◽  
Kathryn Sandberg ◽  
Susette C. Mueller
Keyword(s):  
Confocal Microscopy ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Receptor Type ◽  
3D Analysis ◽  
Kidney Cells ◽  
Angiotensin Receptor ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells

scholarly journals Similar uptake but different trafficking and escape routes of reovirus virions and infectious subvirion particles imaged in polarized Madin–Darby canine kidney cells

2013 ◽  
Vol 24 (8) ◽  
pp. 1196-1207 ◽  
Author(s):  
Steeve Boulant ◽  
Megan Stanifer ◽  
Comert Kural ◽  
David K. Cureton ◽  
Ramiro Massol ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Endocytic Pathway ◽  
Cell Entry ◽  
Kidney Cells ◽  
Apical Surface ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

Polarized epithelial cells that line the digestive, respiratory, and genitourinary tracts form a barrier that many viruses must breach to infect their hosts. Current understanding of cell entry by mammalian reovirus (MRV) virions and infectious subvirion particles (ISVPs), generated from MRV virions by extracellular proteolysis in the digestive tract, are mostly derived from in vitro studies with nonpolarized cells. Recent live-cell imaging advances allow us for the first time to visualize events at the apical surface of polarized cells. In this study, we used spinning-disk confocal fluorescence microscopy with high temporal and spatial resolution to follow the uptake and trafficking dynamics of single MRV virions and ISVPs at the apical surface of live polarized Madin–Darby canine kidney cells. Both types of particles were internalized by clathrin-mediated endocytosis, but virions and ISVPs exhibited strikingly different trafficking after uptake. While virions reached early and late endosomes, ISVPs did not and instead escaped the endocytic pathway from an earlier location. This study highlights the broad advantages of using live-cell imaging combined with single-particle tracking for identifying key steps in cell entry by viruses.

Continuous live cell imaging of cellulose attachment by microbes under anaerobic and thermophilic conditions using confocal microscopy

2013 ◽  
Vol 25 (5) ◽  
pp. 849-856 ◽  
Author(s):  
Zhi-Wu Wang ◽  
Seung-Hwan Lee ◽  
James G. Elkins ◽  
Yongchao Li ◽  
Scott Hamilton-Brehm ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Thermophilic Conditions

scholarly journals KV1.5–KVβ1.3 Recycling Is PKC-Dependent

2021 ◽  
Vol 22 (3) ◽  
pp. 1336
Author(s):  
Alvaro Macias ◽  
Alicia de la Cruz ◽  
Diego A. Peraza ◽  
Angela de Benito-Bueno ◽  
Teresa Gonzalez ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Hek293 Cells ◽  
Dependent Manner ◽  
Fast Inactivation ◽  
Patch Clamp Technique ◽  
Relevant Issue ◽  
Regulatory Subunits

KV1.5 channel function is modified by different regulatory subunits. KVβ1.3 subunits assemble with KV1.5 channels and induce a fast and incomplete inactivation. Inhibition of PKC abolishes the KVβ1.3-induced fast inactivation, decreases the amplitude of the current KV1.5–KVβ1.3 and modifies their pharmacology likely due to changes in the traffic of KV1.5–KVβ1.3 channels in a PKC-dependent manner. In order to analyze this hypothesis, HEK293 cells were transfected with KV1.5–KVβ1.3 channels, and currents were recorded by whole-cell configuration of the patch-clamp technique. The presence of KV1.5 in the membrane was analyzed by biotinylation techniques, live cell imaging and confocal microscopy approaches. PKC inhibition resulted in a decrease of 33 ± 7% of channels in the cell surface due to reduced recycling to the plasma membrane, as was confirmed by confocal microscopy. Live cell imaging indicated that PKC inhibition almost abolished the recycling of the KV1.5–KVβ1.3 channels, generating an accumulation of channels into the cytoplasm. All these results suggest that the trafficking regulation of KV1.5–KVβ1.3 channels is dependent on phosphorylation by PKC and, therefore, they could represent a clinically relevant issue, mainly in those diseases that exhibit modifications in PKC activity.

Live Cell Imaging and Confocal Microscopy

2018 ◽  
pp. 117-130 ◽  
Author(s):  
Luciana Renna ◽  
Giovanni Stefano ◽  
Federica Brandizzi
Keyword(s):  
Confocal Microscopy ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell
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