Live Cell Imaging Confocal Microscopy Analysis of HBV Myr-PreS1 Peptide Binding and Uptake in NTCP-GFP Expressing HepG2 Cells

2016 ◽  
pp. 27-36 ◽  
Author(s):  
Alexander König ◽  
Dieter Glebe
Keyword(s):  
Confocal Microscopy ◽  
Hepg2 Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Peptide Binding ◽  
Live Cell ◽  
Microscopy Analysis ◽  
Confocal Microscopy Analysis

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

scholarly journals Live cell imaging of cytosolic NADH/NAD+ ratio in hepatocytes and liver slices

2018 ◽  
Vol 314 (1) ◽  
pp. G97-G108 ◽  
Author(s):  
Ricard Masia ◽  
William J. McCarty ◽  
Carolina Lahmann ◽  
Jay Luther ◽  
Raymond T. Chung ◽  
...  
Keyword(s):  
Liver Disease ◽  
Real Time ◽  
Hepg2 Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
The United States ◽  
Live Cell ◽  
New Approach ◽  
Liver Slices ◽  
Metabolic Zonation

Fatty liver disease (FLD), the most common chronic liver disease in the United States, may be caused by alcohol or the metabolic syndrome. Alcohol is oxidized in the cytosol of hepatocytes by alcohol dehydrogenase (ADH), which generates NADH and increases cytosolic NADH/NAD+ ratio. The increased ratio may be important for development of FLD, but our ability to examine this question is hindered by methodological limitations. To address this, we used the genetically encoded fluorescent sensor Peredox to obtain dynamic, real-time measurements of cytosolic NADH/NAD+ ratio in living hepatocytes. Peredox was expressed in dissociated rat hepatocytes and HepG2 cells by transfection, and in mouse liver slices by tail-vein injection of adeno-associated virus (AAV)-encoded sensor. Under control conditions, hepatocytes and liver slices exhibit a relatively low (oxidized) cytosolic NADH/NAD+ ratio as reported by Peredox. The ratio responds rapidly and reversibly to substrates of lactate dehydrogenase (LDH) and sorbitol dehydrogenase (SDH). Ethanol causes a robust dose-dependent increase in cytosolic NADH/NAD+ ratio, and this increase is mitigated by the presence of NAD+-generating substrates of LDH or SDH. In contrast to hepatocytes and slices, HepG2 cells exhibit a relatively high (reduced) ratio and show minimal responses to substrates of ADH and SDH. In slices, we show that comparable results are obtained with epifluorescence imaging and two-photon fluorescence lifetime imaging (2p-FLIM). Live cell imaging with Peredox is a promising new approach to investigate cytosolic NADH/NAD+ ratio in hepatocytes. Imaging in liver slices is particularly attractive because it allows preservation of liver microanatomy and metabolic zonation of hepatocytes. NEW & NOTEWORTHY We describe and validate a new approach for measuring free cytosolic NADH/NAD+ ratio in hepatocytes and liver slices: live cell imaging with the fluorescent biosensor Peredox. This approach yields dynamic, real-time measurements of the ratio in living, functioning liver cells, overcoming many limitations of previous methods for measuring this important redox parameter. The feasibility of using Peredox in liver slices is particularly attractive because slices allow preservation of hepatic microanatomy and metabolic zonation of hepatocytes.

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