Plasma Protein Binding of Luciferase Substrates Influences Sensitivity and Accuracy of Bioluminescence Imaging

Keyaerts, Marleen; Heneweer, Carola; Gainkam, Lea Olive Tchouate; Caveliers, Vicky; Beattie, Bradley J.; Martens, Geert A.; Vanhove, Christian; Bossuyt, Axel; Blasberg, Ronald G.; Lahoutte, Tony

doi:10.1007/s11307-010-0325-x

Cited by 15 publications

(14 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, protein A did not show the luminescence activity and the initial maximum intensity of luminescence ( I max ) was same to that of the background level from coelenterazine (Table ). BSA had very weak luminescence activity with coelenterazine as previously reported , but was only 0.9% of I max to ZZ‐gCys. The I max value of ZZ‐gCys was 2.7 times higher than that of Z‐gCys.…”

Section: Resultssupporting

confidence: 77%

A Novel Catalytic Function of Synthetic IgG‐Binding Domain (Z Domain) from Staphylococcal Protein A: Light Emission with Coelenterazine

Inouye

Sahara-Miura

2013

Photochem & Photobiology

View full text Add to dashboard Cite

The synthetic IgG-binding domain (Z domain) of staphylococcal protein A catalyzes the oxidation of coelenterazine to emit light like a coelenterazine-utilizing luciferase. The Z domain derivatives (ZZ-gCys, Z-gCys and Z-domain) were purified and the luminescence properties were characterized by comparing with coelenterazine-utilizing luciferases, including Renilla luciferase, Gaussia luciferase and the catalytic 19 kDa protein of Oplophorus luciferase. Three Z domain derivatives showed luminescence activity with coelenterazine and the order of the initial maximum intensity of luminescence was ZZ-gCys (100%) > Z-gCys (36.8%) > Z-domain (1.1%) > bovine serum albumin (BSA; 0.9%) > staphylococcal protein A (0.1%) and the background value of coelenterazine (0.1%) in our conditions. The luminescence properties of ZZ-gCys showed the similarity to that of Gaussia luciferase, including the luminescence pattern, the emission spectrum, the stimulation by halogen ions and nonionic detergents and the substrate specificity for coelenterazine analogues. In contrast, the luminescence properties of Z-gCys were close to the catalytic 19 kDa protein of Oplophorus luciferase. The catalytic region of the Z domain for the luminescence reaction might be different from the IgG-binding region of the Z domain.

show abstract

Section: Resultssupporting

confidence: 77%

A Novel Catalytic Function of Synthetic IgG‐Binding Domain (Z Domain) from Staphylococcal Protein A: Light Emission with Coelenterazine

Inouye

Sahara-Miura

2013

Photochem & Photobiology

View full text Add to dashboard Cite

show abstract

“…In vitro , a linear correlation exists between PE and number of luciferase expressing cells. In vivo this relation is affected by luciferin distribution and photon absorption and scattering by tissue (Virostko et al, 2004; Inoue et al, 2009; Keyaerts et al, 2011). Bioluminence imaging of the brain has to deal with limited substrate diffusibility through the blood brain-barrier (BBB; Berger et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

In vivo bioluminescence imaging of vascular remodeling after stroke

Adamczak

Schneider

Nelles

et al. 2014

Front. Cell. Neurosci.

View full text Add to dashboard Cite

Thrombolysis remains the only beneficial therapy for ischemic stroke, but is restricted to a short therapeutic window following the infarct. Currently research is focusing on spontaneous regenerative processes during the sub-acute and chronic phase. Angiogenesis, the formation of new blood vessels from pre-existing ones, was observed in stroke patients, correlates with longer survival and positively affects the formation of new neurons. Angiogenesis takes place in the border zones of the infarct, but further insight into the temporal profile is needed to fully apprehend its therapeutic potential and its relevance for neurogenesis and functional recovery. Angiogenesis is a multistep process, involving extracellular matrix degradation, endothelial cell proliferation, and, finally, new vessel formation. Interaction between vascular endothelial growth factor and its receptor 2 (VEGFR2) plays a central role in these angiogenic signaling cascades. In the present study we investigated non-invasively the dynamics of VEGFR2 expression following cerebral ischemia in a mouse model of middle cerebral artery occlusion (MCAO). We used a transgenic mouse expressing firefly luciferase under the control of the VEGFR2 promotor to non-invasively elucidate the temporal profile of VEGFR2 expression after stroke as a biomarker for VEGF/VEGFR2 signaling. We measured each animal repetitively up to 2 weeks after stroke and found increased VEGFR2 expression starting 3 days after the insult with peak values at 7 days. These were paralleled by increased VEGFR2 protein levels and increased vascular volume in peri-infarct areas at 14 days after the infarct, indicating that signaling via VEGFR2 leads to successful vascular remodeling. This study describes VEGFR2-related signaling is active at least up to 2 weeks after the infarct and results in increased vascular volume. Further, this study presents a novel strategy for the non-invasive evaluation of angiogenesis-based therapies.

show abstract

“…Mice were imaged at day 3, 5 and 7 post cell injection, alternating between isoflurane anesthesia (2%) and the test anesthetic with a cross over design, so that half of the mice had twice isoflurane and half of the mice had twice the test anesthetic [3]. For the final signal intensity, the average of day 3 and day 7 was compared to day 5, to correct for cell growth between the different imaging sessions.…”

Section: Methodsmentioning

confidence: 99%

“…The signal intensity reflects the strength or changes of a molecular target in a quantitative way. In vivo , many other parameters can influence the BLI readout, such as the administration route and protein binding of the substrates, overlying tissues characteristics and membrane pumps that can remove substrates out of the cell [1], [2], [3], [4], [5], [6].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2012

Self Cite

View full text Add to dashboard Cite

Bioluminescence imaging is routinely performed in anesthetized mice. Often isoflurane anesthesia is used because of its ease of use and fast induction/recovery. However, general anesthetics have been described as important inhibitors of the luciferase enzyme reaction.AimTo investigate frequently used mouse anesthetics for their direct effect on the luciferase reaction, both in vitro and in vivo.Materials and Methodsisoflurane, sevoflurane, desflurane, ketamine, xylazine, medetomidine, pentobarbital and avertin were tested in vitro on luciferase-expressing intact cells, and for non-volatile anesthetics on intact cells and cell lysates. In vivo, isoflurane was compared to unanesthetized animals and different anesthetics. Differences in maximal photon emission and time-to-peak photon emission were analyzed.ResultsAll volatile anesthetics showed a clear inhibitory effect on the luciferase activity of 50% at physiological concentrations. Avertin had a stronger inhibitory effect of 80%. For ketamine and xylazine, increased photon emission was observed in intact cells, but this was not present in cell lysate assays, and was most likely due to cell toxicity and increased cell membrane permeability. In vivo, the highest signal intensities were measured in unanesthetized mice and pentobarbital anesthetized mice, followed by avertin. Isoflurane and ketamine/medetomidine anesthetized mice showed the lowest photon emission (40% of unanesthetized), with significantly longer time-to-peak than unanesthetized, pentobarbital or avertin-anesthetized mice. We conclude that, although strong inhibitory effects of anesthetics are present in vitro, their effect on in vivo BLI quantification is mainly due to their hemodynamic effects on mice and only to a lesser extent due to the direct inhibitory effect.

show abstract

Plasma Protein Binding of Luciferase Substrates Influences Sensitivity and Accuracy of Bioluminescence Imaging

Cited by 15 publications

References 20 publications

A Novel Catalytic Function of Synthetic IgG‐Binding Domain (Z Domain) from Staphylococcal Protein A: Light Emission with Coelenterazine

A Novel Catalytic Function of Synthetic IgG‐Binding Domain (Z Domain) from Staphylococcal Protein A: Light Emission with Coelenterazine

In vivo bioluminescence imaging of vascular remodeling after stroke

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

Contact Info

Product

Resources

About