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Methods for Detecting PER2:LUCIFERASE Bioluminescence Rhythms in Freely Moving Mice.

Blanca Martin-Burgos1, Wanqi Wang1, Ivana William1

  • 1Neuroscience Program, Smith College, Northampton, Massachusetts.

Journal of Biological Rhythms
|December 7, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a method to measure circadian rhythms in freely moving mice using bioluminescence. This technique allows for the study of peripheral clocks and their responses to various stimuli.

Keywords:
CycLuc1PERIOD2bioluminescencecircadianin vivoluciferaseperipheral oscillatorsreporter gene

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Area of Science:

  • Chronobiology
  • Molecular Biology
  • Animal Models

Background:

  • Circadian rhythms are daily gene expression oscillations.
  • Bioluminescent reporters, like luciferase, are crucial for studying these rhythms in cells and tissues.
  • Measuring these rhythms in freely moving animals presents technical challenges.

Purpose of the Study:

  • To establish a method for measuring circadian bioluminescence in freely moving mice.
  • To optimize conditions for detecting circadian rhythms using the PER2::LUC reporter.
  • To evaluate different substrate delivery methods and identify suitable substrates for long-term studies.

Main Methods:

  • Utilized the LumiCycle In Vivo system for bioluminescence measurement in standard cages.
  • Tested subcutaneous minipump and drinking water delivery for d-luciferin.
  • Employed the synthetic luciferase substrate CycLuc1 for extended monitoring.
  • Analyzed bioluminescence rhythms, including sex-based phase differences.

Main Results:

  • Successfully detected circadian rhythms of PER2::LUC bioluminescence in freely moving mice.
  • Observed bioluminescence spikes correlated with drinking bouts.
  • Demonstrated that CycLuc1 supports circadian rhythms at lower concentrations and for longer durations than d-luciferin.
  • Identified a phase advance in female mice compared to males.

Conclusions:

  • The described method enables robust measurement of circadian rhythms in freely moving mice.
  • This technique is adaptable for tissue-specific reporters, facilitating the study of peripheral clocks.
  • The findings provide a foundation for investigating how environmental and pharmacological factors impact circadian rhythms in specific tissues.