Coelenterazine (Synonyms: Luciferin (Oplophorus), Preluciferin (Watasenia))

Cell lines

RAW 264.7 cells

Preparation Method

5 × 10⁴ cRAW 264.7 cells were added to wells of a 96-well black plate in triplicate, cell counts were performed, and native coelenterazine was added to the sample (final coelenterazine concentration 0-100 μmol/l). Plates were imaged using an IVIS low-light imaging system

Reaction Conditions

final coelenterazine concentration 0-100 μmol/l，60min

Applications

RAW 264.7 chemiluminescent signal intensity positively correlates with coelenterazine concentration in culture. The production of chemiluminescence by RAW 264.7 cells in the absence of stimulation is consistent with the detection of superoxide anion production during oxidative phosphorylation.

Animal models

10-26 weeks thirty-six mice (16 females, 20 males)

Preparation Method

450 nl of virus (three sites) were injected into the cerebral cortex of mice, followed by coelenterazine injections at 28-day intervals (minimum = 26 days, maximum time = 74 days). Coelenterazine (dissolved in DMSO) and diluted to a concentration of 2.43 mM in sterile water (1 mg/ml) for cortical injection through a 34-gauge needle for imaging and electrophysiological recordings.

Dosage form

0.2µl, 0.4µl and 1µl，the injection rate was 1.25 μL/min

Applications

Coelenterazine strongly boosted activity in LMO3-expressing mice, photon production was proportional to emission rate, and maximal doses of coelenterazine moderately increased neural activity in naive mice and GLuc cohorts, which itself can be produced at high concentrations Nonspecific excitatory effects.

References:

[1]. Bronsart L, Nguyen L, Habtezion A, Contag C. Reactive Oxygen Species Imaging in a Mouse Model of Inflammatory Bowel Disease. Mol Imaging Biol. 2016 Aug;18(4):473-8.

[2]. Gomez-Ramirez M, More AI,et al. The BioLuminescent-OptoGenetic in vivo response to coelenterazine is proportional, sensitive, and specific in neocortex. J Neurosci Res. 2020 Mar;98(3):471-480.