scholarly journals Red-Shifted Firefly Luciferase Optimized for Candida albicans In vivo Bioluminescence Imaging

2017 ◽  
Vol 8 ◽  
Author(s):  
Stephane Dorsaz ◽  
Alix T. Coste ◽  
Dominique Sanglard
Keyword(s):  
Candida Albicans ◽  
Bioluminescence Imaging ◽  
Firefly Luciferase ◽  
In Vivo Bioluminescence Imaging

scholarly journals Sensitive Dual Color In Vivo Bioluminescence Imaging Using a New Red Codon Optimized Firefly Luciferase and a Green Click Beetle Luciferase

PLoS ONE ◽  
2011 ◽  
Vol 6 (4) ◽  
pp. e19277 ◽  
Author(s):  
Laura Mezzanotte ◽  
Ivo Que ◽  
Eric Kaijzel ◽  
Bruce Branchini ◽  
Aldo Roda ◽  
...  
Keyword(s):  
Bioluminescence Imaging ◽  
Firefly Luciferase ◽  
Dual Color ◽  
Click Beetle ◽  
In Vivo Bioluminescence Imaging

scholarly journals A synthetic luciferin improves in vivo bioluminescence imaging of gene expression in cardiovascular brain regions

2016 ◽  
Vol 48 (10) ◽  
pp. 762-770 ◽  
Author(s):  
Hayk Simonyan ◽  
Chansol Hurr ◽  
Colin N. Young
Keyword(s):  
Gene Expression ◽  
Targeted Delivery ◽  
Bioluminescence Imaging ◽  
Firefly Luciferase ◽  
Brain Regions ◽  
Cmv Promoter ◽  
Brain Nuclei ◽  
Molecular Events ◽  
In Vivo Bioluminescence Imaging

Bioluminescence imaging is an effective tool for in vivo investigation of molecular processes. We have demonstrated the applicability of bioluminescence imaging to spatiotemporally monitor gene expression in cardioregulatory brain nuclei during the development of cardiovascular disease, via incorporation of firefly luciferase into living animals, combined with exogenous d-luciferin substrate administration. Nevertheless, d-luciferin uptake into the brain tissue is low, which decreases the sensitivity of bioluminescence detection, particularly when considering small changes in gene expression in tiny central areas. Here, we tested the hypothesis that a synthetic luciferin, cyclic alkylaminoluciferin (CycLuc1), would be superior to d-luciferin for in vivo bioluminescence imaging in cardiovascular brain regions. Male C57B1/6 mice underwent targeted delivery of an adenovirus encoding the luciferase gene downstream of the CMV promoter to the subfornical organ (SFO) or paraventricular nucleus of hypothalamus (PVN), two crucial cardioregulatory neural regions. While bioluminescent signals could be obtained following d-luciferin injection (150 mg/kg), CycLuc1 administration resulted in a three- to fourfold greater bioluminescent emission from the SFO and PVN, at 10- to 20-fold lower substrate concentrations (7.5–15 mg/kg). This CycLuc1-mediated enhancement in bioluminescent emission was evident early following substrate administration (i.e., 6–10 min) and persisted for up to 1 h. When the exposure time was reduced from 60 s to 1,500 ms, minimal signal in the PVN was detectable with d-luciferin, whereas bioluminescent images could be reliably captured with CycLuc1. These findings demonstrate that bioluminescent imaging with the synthetic luciferin CycLuc1 provides an improved physiological genomics tool to investigate molecular events in discrete cardioregulatory brain nuclei.

Abstract MP04: In Vivo Bioluminescence Imaging Reveals a Pre-Hypertensive Surge of Nuclear-Factor-κ-B (NF-κB) Activity in the Subfornical Organ (SFO) During Obesity-Induced Hypertension

Hypertension ◽  
2014 ◽  
Vol 64 (suppl_1) ◽  
Author(s):  
Colin N Young ◽  
Scott D Butler ◽  
Robin L Davisson
Keyword(s):  
Bioluminescence Imaging ◽  
Firefly Luciferase ◽  
Induced Hypertension ◽  
In Vivo Bioluminescence Imaging ◽  
Normal Chow ◽  
Brain Center ◽  
The Central Nervous System ◽  
Underlying Mechanisms ◽  
Nuclear Factor Κ B

The central nervous system has been implicated in obesity-induced sympathetic activation and hypertension, although the underlying mechanisms remain unclear. We tested the hypothesis that the transcription factor NF-κB is activated in the SFO, a key cardioregulatory brain center, during the development of obesity-induced hypertension. Male C57Bl/6 mice were instrumented with radiotelemeters for mean arterial pressure (MAP) recording, and underwent SFO-targeted microinjection of an adenovirus encoding firefly luciferase downstream of the NF-κB response elements. After recovery, mice were fed high fat diet (60% fat) or normal chow (5% fat), and SFO NF-κB activity was longitudinally monitored in vivo by bioluminescence imaging coupled with luciferin injections. In response to HFD, a slowly developing hypertension was observed, with an initial rise in MAP at 4 weeks (109 ± 1 vs 117 ± 1 mmHg; day 0 vs day 28; n=8; p<0.05) that was sustained for 8 weeks. NF-κB activity in the SFO increased gradually during HFD feeding, with a surge at 3 weeks, prior to a rise in MAP (day 21: 0.91 ± 0.08 vs 1.70 ± 0.25 photons/s fold baseline; normal chow vs HFD; n=4-7; p<0.05). SFO NF-κB activity transiently subsided and a second elevation was noted during the maintenance of obesity-induced hypertension at 6.5 weeks (day 42: 0.72 ± 0.14 vs 1.52 ± 0.07 photons/s fold baseline; normal chow vs HFD; n=4-7; p<0.05). These findings suggest that NF-κB is activated in the SFO during HFD feeding. The biphasic increase in NF-κB activity prior to and during a sustained increase in MAP suggests a casual role for SFO NF-κB in the development of obesity-induced hypertension. HL63887, HL84207, AHA13POST14410020, K99HL166776

scholarly journals Inhibition of Firefly Luciferase by General Anesthetics: Effect on In Vitro and In Vivo Bioluminescence Imaging

PLoS ONE ◽  
2012 ◽  
Vol 7 (1) ◽  
pp. e30061 ◽  
Author(s):  
Marleen Keyaerts ◽  
Isabel Remory ◽  
Vicky Caveliers ◽  
Karine Breckpot ◽  
Tomas J. Bos ◽  
...  
Keyword(s):  
Bioluminescence Imaging ◽  
Firefly Luciferase ◽  
General Anesthetics ◽  
In Vivo Bioluminescence Imaging

scholarly journals Multicolour In Vivo Bioluminescence Imaging Using a NanoLuc‐Based BRET Reporter in Combination with Firefly Luciferase

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Arthur Taylor ◽  
Jack Sharkey ◽  
Antonius Plagge ◽  
Bettina Wilm ◽  
Patricia Murray
Keyword(s):  
Bioluminescence Imaging ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
High Sensitivity ◽  
Firefly Luciferase ◽  
Cell Populations ◽  
Wide Range ◽  
In Vivo Bioluminescence Imaging ◽  
Multiple Cell

The ability to track the biodistribution and fate of multiple cell populations administered to rodents has the potential to facilitate the understanding of biological processes in a range of fields including regenerative medicine, oncology, and host/pathogen interactions. Bioluminescence imaging is an important tool for achieving this goal, but current protocols rely on systems that have poor sensitivity or require spectral decomposition. Here, we show that a bioluminescence resonance energy transfer reporter (BRET) based on NanoLuc and LSSmOrange in combination with firefly luciferase enables the unambiguous discrimination of two cell populations in vivo with high sensitivity. We insert each of these reporter genes into cells using lentiviral vectors and demonstrate the ability to monitor the cells’ biodistribution under a wide range of administration conditions, including the venous or arterial route, and in different tissues including the brain, liver, kidneys, and tumours. Our protocol allows for the imaging of two cell populations in the same imaging session, facilitating the overlay of the signals and the identification of anatomical positions where they colocalise. Finally, we provide a method for postmortem confirmation of the presence of each cell population in excised organs.

The Antitumor Effects of Britanin on Hepatocellular Carcinoma Cells and its Real-Time Evaluation by In Vivo Bioluminescence Imaging

2020 ◽  
Vol 20 (9) ◽  
pp. 1147-1156
Author(s):  
Hanrui Li ◽  
GeTao Du ◽  
Lu Yang ◽  
Liaojun Pang ◽  
Yonghua Zhan
Keyword(s):  
Hepatocellular Carcinoma ◽  
Western Blot ◽  
Blot Analysis ◽  
Tumor Size ◽  
Hepg2 Cells ◽  
Western Blot Analysis ◽  
Bioluminescence Imaging ◽  
Antitumor Effects ◽  
In Vivo Bioluminescence Imaging

Background: Hepatocellular carcinoma is cancer with many new cases and the highest mortality rate. Chemotherapy is the most commonly used method for the clinical treatment of hepatocellular carcinoma. Natural products have become clinically important chemotherapeutic drugs due to their great potential for pharmacological development. Many sesquiterpene lactone compounds have been proven to have antitumor effects on hepatocellular carcinoma. Objective: Britanin is a sesquiterpene lactone compound that can be considered for the treatment of hepatocellular carcinoma. The present study aimed to investigate the antitumor effect of britanin. Methods: BEL 7402 and HepG2 cells were used to study the cytotoxicity and antitumor effects of britanin. Preliminary studies on the nuclear factor kappa B pathway were conducted by western blot analysis. A BEL 7402-luc subcutaneous tumor model was established for the in vivo antitumor studies of britanin. In vivo bioluminescence imaging was conducted to monitor changes in tumor size. Results: The results of the cytotoxicity analysis showed that the IC50 values for britanin in BEL 7402 and HepG2 cells were 2.702μM and 6.006μM, respectively. The results of the colony formation demonstrated that the number of cells in a colony was reduced significantly after britanin treatment. And the results of transwell migration assays showed that the migration ability of tumor cells was significantly weakened after treatment with britanin. Tumor size measurements and staining results showed that tumor size was inhibited after britanin treatment. The western blot analysis results showed the inhibition of p65 protein expression and reduced the ratio of Bcl-2/Bax after treatment. Conclusion: A series of in vitro and in vivo experiments demonstrated that britanin had good antitumor effects and provided an option for hepatocellular carcinoma treatment.

In Vivo Bioluminescence Imaging of Transplanted Islets and Early Detection of Graft Rejection

2006 ◽  
Vol 81 (10) ◽  
pp. 1421-1427 ◽  
Author(s):  
Xiaojuan Chen ◽  
Xiaomin Zhang ◽  
Courtney S. Larson ◽  
Marshall S. Baker ◽  
Dixon B. Kaufman
Keyword(s):  
Early Detection ◽  
Graft Rejection ◽  
Bioluminescence Imaging ◽  
In Vivo Bioluminescence Imaging
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