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Using bioluminescence to image gene expression and spontaneous behavior in freely moving mice
Astha Malik1, Jessica A Zavadil2, Michael E Geusz3
1Division of Gastroenterology, Hepatology, & Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
Plos One
|January 20, 2023
Summary
This study presents a novel bioluminescence imaging (BLI) technique for live animals, enabling long-term gene expression monitoring in freely moving, unanesthetized subjects. The method reduces stress and improves data accuracy for biological process studies.
Area of Science:
- Live animal imaging
- Gene expression analysis
- Bioluminescence imaging (BLI)
Background:
- Bioluminescence imaging (BLI) is crucial for monitoring dynamic biological processes in live animals.
- Conventional BLI methods often involve anesthesia and stressors, potentially altering results and limiting long-term studies.
- Existing techniques struggle with rhythmic gene expression effects and animal movement artifacts.
Purpose of the Study:
- To develop a minimally stressful BLI strategy for sustained gene expression monitoring in freely moving, unanesthetized mice.
- To overcome limitations of conventional BLI, including circadian rhythm interference and physiological alterations due to stress.
- To enable accurate, long-term tracking of gene activity in awake, behaving animals.
Main Methods:
- Developed a novel BLI approach minimizing animal stress before and during imaging.
- Utilized hairless albino mice expressing luciferase, allowing luciferin ingestion during normal behavior.
- Employed cooled electron-multiplying-CCD cameras and computer-aided image selection for consistent posture-based measurements.
- Performed imaging primarily during the animal's night, compatible with circadian entrainment and multi-day sampling.
Main Results:
- Successfully monitored Per1 gene expression, a component of the circadian clock, in freely moving, unanesthetized mice.
- Demonstrated the technique's effectiveness in minimizing stress-induced physiological and behavioral alterations.
- Achieved comparable measurements over long intervals by standardizing imaging posture.
- Showed compatibility with circadian entrainment and enabled multi-day sampling for long-term change monitoring.
Conclusions:
- The developed BLI strategy offers an effective alternative to imaging immobile, anesthetized animals.
- This method reduces noise from circadian oscillations and body movements, enhancing data quality for long-term studies.
- Minimizing stress and allowing natural behavior improves the reliability of gene expression monitoring in live animals.
