Video rate bioluminescence imaging of secretory proteins in living cells: Localization, secretory frequency, and quantification

2011 ◽  
Vol 415 (2) ◽  
pp. 182-189 ◽  
Author(s):  
Takahiro Suzuki ◽  
Chihiro Kondo ◽  
Takao Kanamori ◽  
Satoshi Inouye
Keyword(s):  
Bioluminescence Imaging ◽  
Living Cells ◽  
Secretory Proteins

A caged 2‐hydroxyethyl luciferin for bioluminescence imaging of nitroxyl in living cells

Luminescence ◽  
2020 ◽  
Vol 35 (8) ◽  
pp. 1384-1390
Author(s):  
Yong Ju ◽  
Anni Wang ◽  
Xuewei Li ◽  
Xu Xu ◽  
Jianzhong Lu
Keyword(s):  
Bioluminescence Imaging ◽  
Living Cells

scholarly journals Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system

2019 ◽  
Vol 116 (52) ◽  
pp. 26491-26496 ◽  
Author(s):  
Carola Gregor ◽  
Jasmin K. Pape ◽  
Klaus C. Gwosch ◽  
Tanja Gilat ◽  
Steffen J. Sahl ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Bioluminescence Imaging ◽  
Single Cells ◽  
Cell Types ◽  
Firefly Luciferase ◽  
Living Cells ◽  
Spatiotemporal Resolution ◽  
Bacterial Bioluminescence ◽  
Mammalian Cell Lines ◽  
Bioluminescence Signal

Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterialluxCDABEandfrpgenes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells.

Bioluminescence imaging of carbon monoxide in living cells based on a selective deiodination reaction

The Analyst ◽  
2020 ◽  
Vol 145 (2) ◽  
pp. 550-556 ◽  
Author(s):  
Anni Wang ◽  
Xuewei Li ◽  
Yong Ju ◽  
Dongying Chen ◽  
Jianzhong Lu
Keyword(s):  
Carbon Monoxide ◽  
Cancer Cells ◽  
Iodine Atom ◽  
Bioluminescence Imaging ◽  
Living Cells ◽  
Turn On ◽  
Co Detection

Modification of a heavy iodine atom for d-Luciferin was explored as a “turn-on” transduction scheme for CO detection. This new probe could image exogenous and endogenous CO in the luciferase-transfected cancer cells.

scholarly journals Kar2p availability defines distinct forms of endoplasmic reticulum stress in living cells

2012 ◽  
Vol 23 (5) ◽  
pp. 955-964 ◽  
Author(s):  
Patrick Lajoie ◽  
Robyn D. Moir ◽  
Ian M. Willis ◽  
Erik L. Snapp
Keyword(s):  
Endoplasmic Reticulum ◽  
Fluorescent Protein ◽  
Secretory Protein ◽  
Living Cells ◽  
Secretory Proteins ◽  
High Throughput Analysis ◽  
Protein Levels ◽  
Unfolded Protein ◽  
Green Fluorescent ◽  
The Unfolded Protein Response

Accumulation of misfolded secretory proteins in the endoplasmic reticulum (ER) activates the unfolded protein response (UPR) stress pathway. To enhance secretory protein folding and promote adaptation to stress, the UPR upregulates ER chaperone levels, including BiP. Here we describe chromosomal tagging of KAR2, the yeast homologue of BiP, with superfolder green fluorescent protein (sfGFP) to create a multifunctional endogenous reporter of the ER folding environment. Changes in Kar2p-sfGFP fluorescence levels directly correlate with UPR activity and represent a robust reporter for high-throughput analysis. A novel second feature of this reporter is that photobleaching microscopy (fluorescence recovery after photobleaching) of Kar2p-sfGFP mobility reports on the levels of unfolded secretory proteins in individual cells, independent of UPR status. Kar2p-sfGFP mobility decreases upon treatment with tunicamycin or dithiothreitol, consistent with increased levels of unfolded proteins and the incorporation of Kar2p-sfGFP into slower-diffusing complexes. During adaptation, we observe a significant lag between down-regulation of the UPR and resolution of the unfolded protein burden. Finally, we find that Kar2p-sfGFP mobility significantly increases upon inositol withdrawal, which also activates the UPR, apparently independent of unfolded protein levels. Thus Kar2p mobility represents a powerful new tool capable of distinguishing between the different mechanisms leading to UPR activation in living cells.

scholarly journals Prolonged bioluminescence imaging in living cells and mice using novel pro-substrates forRenillaluciferase

2017 ◽  
Vol 15 (48) ◽  
pp. 10238-10244 ◽  
Author(s):  
Mingliang Yuan ◽  
Xiaojie Ma ◽  
Tianyu Jiang ◽  
Yuqi Gao ◽  
Yuanyuan Cui ◽  
...  
Keyword(s):  
Bioluminescence Imaging ◽  
Living Cells

The prodrug or caged-luciferin strategy affords an excellent platform for persistent bioluminescence imaging.

Bioluminescence Imaging of Carbon Monoxide in Living Cells and Nude Mice Based on Pd0-Mediated Tsuji–Trost Reaction

2018 ◽  
Vol 90 (9) ◽  
pp. 5951-5958 ◽  
Author(s):  
Xiaodong Tian ◽  
Xinda Liu ◽  
Anni Wang ◽  
Choiwan Lau ◽  
Jianzhong Lu
Keyword(s):  
Carbon Monoxide ◽  
Nude Mice ◽  
Bioluminescence Imaging ◽  
Living Cells

Ultrastructural Aspects of Golgi Complex Function in Parathyroid Cells of Winter Frogs

1973 ◽  
Vol 31 ◽  
pp. 680-681
Author(s):  
William J. Dougherty
Keyword(s):  
Golgi Complex ◽  
Secretory Granules ◽  
Complex Function ◽  
Secretory Activity ◽  
Secretory Proteins ◽  
Perivascular Spaces ◽  
Pituitary Cells ◽  
Electron Microscopic ◽  
Ca Ions ◽  
Regulation Of Secretion

The regulation of secretion in exocrine and endocrine cells has long been of interest. Electron microscopic and other studies have demonstrated that secretory proteins synthesized on ribosomes are transported by the rough ER to the Golgi complex where they are concentrated into secretory granules. During active secretion, secretory granules fuse with the cell membrane, liberating and discharging their contents into the perivascular spaces. When secretory activity is suppressed in anterior pituitary cells, undischarged secretory granules may be degraded by lysosomes. In the parathyroid gland, evidence indicates that the level of blood Ca ions regulates both the production and release of parathormone. Thus, when serum Ca is low, synthesis and release of parathormone are both stimulated; when serum Ca is elevated, these processes are inhibited.

Fluorescence ratio imaging of dynamic intracellular ionic signals

1988 ◽  
Vol 46 ◽  
pp. 42-43
Author(s):  
R. Y. Tsien ◽  
A. Minta ◽  
M. Poenie ◽  
J.P.Y. Kao ◽  
A. Harootunian
Keyword(s):  
Binding Sites ◽  
Living Cells ◽  
Molecular Engineering ◽  
Ph Indicators ◽  
Highly Sensitive ◽  
Fluorescence Ratio Imaging ◽  
Ratio Imaging ◽  
Technical Advances ◽  
Indicator Dyes ◽  
Fluorescein Dyes

Recent technical advances now enable the continuous imaging of important ionic signals inside individual living cells with micron spatial resolution and subsecond time resolution. This methodology relies on the molecular engineering of indicator dyes whose fluorescence is strong and highly sensitive to ions such as Ca2+, H+, or Na+, or Mg2+. The Ca2+ indicators, exemplified by fura-2 and indo-1, derive their high affinity (Kd near 200 nM) and selectivity for Ca2+ to a versatile tetracarboxylate binding site3 modeled on and isosteric with the well known chelator EGTA. The most commonly used pH indicators are fluorescein dyes (such as BCECF) modified to adjust their pKa's and improve their retention inside cells. Na+ indicators are crown ethers with cavity sizes chosen to select Na+ over K+: Mg2+ indicators use tricarboxylate binding sites truncated from those of the Ca2+ chelators, resulting in a more compact arrangement of carboxylates to suit the smaller ion.

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