scholarly journals Live-cell imaging study of mitochondrial morphology in mammalian cells irradiated

2014 ◽  
Vol 55 (suppl 1) ◽  
pp. i129-i130 ◽  
Author(s):  
Y. Kanari ◽  
M. Noguchi ◽  
K. Kaminaga ◽  
Y. Sakamoto ◽  
A. Yokoya
Keyword(s):  
Mammalian Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging Study ◽  
Live Cell ◽  
Mitochondrial Morphology

Live-cell imaging study of mitochondrial morphology in mammalian cells exposed to X-rays

2015 ◽  
Vol 166 (1-4) ◽  
pp. 101-103 ◽  
Author(s):  
M. Noguchi ◽  
Y. Kanari ◽  
A. Yokoya ◽  
A. Narita ◽  
K. Fujii
Keyword(s):  
Mammalian Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging Study ◽  
Live Cell ◽  
Mitochondrial Morphology ◽  
X Rays

The Fate of Nascent APP in Hippocampal Neurons: A Live Cell Imaging Study

2018 ◽  
Vol 9 (9) ◽  
pp. 2225-2232
Author(s):  
Claire E. DelBove ◽  
Xian-zhen Deng ◽  
Qi Zhang
Keyword(s):  
Hippocampal Neurons ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging Study ◽  
Live Cell

scholarly journals Live Cell Imaging of Bioorthogonally Labelled Proteins Generated With a Single Pyrrolysine tRNA Gene

2017 ◽  
Author(s):  
Noa Aloush ◽  
Tomer Schvartz ◽  
Andres I. König ◽  
Sarit Cohen ◽  
Eugene Brozgol ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Stop Codon ◽  
Signal To Noise Ratio ◽  
Trna Synthetase ◽  
Live Cell ◽  
Live Cells ◽  
Copy Numbers ◽  
Intracellular Proteins

ABSTRACTGenetic code expansion enables the incorporation of non-canonical amino acids (ncAAs) into expressed proteins. ncAAs are usually encoded by a stop codon that is decoded by an exogenous orthogonal aminoacyl tRNA synthetase and its cognate suppressor tRNA, such as the pyrrolysine synthetase/ pair. In such systems, stop codon suppression is dependent on the intracellular levels of the exogenous tRNA. Therefore, multiple copies of the tRNAPyl gene (PylT) are encoded to improve ncAA incorporation. However, certain applications in mammalian cells, such as live-cell imaging applications, where labelled tRNA contributes to background fluorescence, can benefit from the use of less invasive minimal expression systems. Accordingly, we studied the effect of tRNAPyl on live-cell fluorescence imaging of bioorthogonally-labelled intracellular proteins. We found that in COS7 cells, a decrease in PylT copy numbers had no measurable effect on protein expression levels. Importantly, reducing PylT copy numbers improved the quality of live-cells images by enhancing the signal-to-noise ratio and reducing an immobile tRNAPyl population. This enabled us to improve live cell imaging of bioorthogonally labelled intracellular proteins, and to simultaneously label two different proteins in a cell. Our results indicate that the number of introduced PylT genes can be minimized according to the transfected cell line, incorporated ncAA, and application.

scholarly journals Sortase-Modified Cholera Toxoids Show Specific Golgi Localization

2019 ◽  
Author(s):  
Darren Machin ◽  
Daniel Williamson ◽  
Peter Fisher ◽  
victoria miller ◽  
Gemma Wildsmith ◽  
...  
Keyword(s):  
Lipid Rafts ◽  
Cell Biology ◽  
Mammalian Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Neuronal Tracing ◽  
Future Role ◽  
Golgi Localization

Cholera toxoid is an established tool for use in cellular tracing in neuroscience and cell biology. We use a sortase-labelling approach to generate site-specifically <i>N</i>-terminally modified variants of both the A2-B<sub>5</sub> heterohexamer and B<sub>5</sub> pentamer forms of the toxoid. Both forms of the toxoid are endocytosed by GM1-positive mammalian cells, and while the heterohexameric toxoid was principally localized in the ER, the B<sub>5</sub> pentamer showed an unexpected localization in the <i>medial/trans</i> Golgi. This study suggests a future role for specifically-labelled cholera toxoids in live-cell imaging beyond their current applications in neuronal tracing and labelling of lipid-rafts in fixed cells.

scholarly journals Cytosolic GPR37, but not GPR37L1, multimerization and its reversal by Parkin: A live cell imaging study

2021 ◽  
Vol 35 (12) ◽  
Author(s):  
Tianyi Li ◽  
Sho Oasa ◽  
Francisco Ciruela ◽  
Lars Terenius ◽  
Vladana Vukojević ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging Study ◽  
Live Cell

scholarly journals Myosin-independent cytokinesis inGiardiautilizes flagella to coordinate force generation and direct membrane trafficking

2017 ◽  
Vol 114 (29) ◽  
pp. E5854-E5863 ◽  
Author(s):  
William R. Hardin ◽  
Renyu Li ◽  
Jason Xu ◽  
Andrew M. Shelton ◽  
Germain C. M. Alas ◽  
...  
Keyword(s):  
Membrane Trafficking ◽  
Mammalian Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Myosin Ii ◽  
Live Cell ◽  
Actin Binding ◽  
Alternative Mechanism ◽  
Membrane Tension ◽  
Contractile Ring

Devoid of all known canonical actin-binding proteins, the prevalent parasiteGiardia lambliauses an alternative mechanism for cytokinesis. Unique aspects of this mechanism can potentially be leveraged for therapeutic development. Here, live-cell imaging methods were developed forGiardiato establish division kinetics and the core division machinery. Surprisingly,Giardiacytokinesis occurred with a median time that is ∼60 times faster than mammalian cells. In contrast to cells that use a contractile ring, actin was not concentrated in the furrow and was not directly required for furrow progression. Live-cell imaging and morpholino depletion of axonemal Paralyzed Flagella 16 indicated that flagella-based forces initiated daughter cell separation and provided a source for membrane tension. Inhibition of membrane partitioning blocked furrow progression, indicating a requirement for membrane trafficking to support furrow advancement. Rab11 was found to load onto the intracytoplasmic axonemes late in mitosis and to accumulate near the ends of nascent axonemes. These developing axonemes were positioned to coordinate trafficking into the furrow and mark the center of the cell in lieu of a midbody/phragmoplast. We show that flagella motility, Rab11, and actin coordination are necessary for proper abscission. Organisms representing three of the five eukaryotic supergroups lack myosin II of the actomyosin contractile ring. These results support an emerging view that flagella play a central role in cell division among protists that lack myosin II and additionally implicate the broad use of membrane tension as a mechanism to drive abscission.

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