Abstract
The activity of the immune system is controlled by circadian clocks present in different immune cells. The brain-resident subtype of immune cells, microglia, exhibits a wide range of functional phenotypes depending on the signaling molecules in their microenvironment. The exact role of microglia in the hypothalamic suprachiasmatic nuclei (SCN), the central circadian clock, has not been known. Therefore, the aim of this study was to determine (1) whether microenvironment-induced changes in microglial polarization affect circadian clocks in these cells and (2) whether the presence of microglia contributes to SCN clock function. Microglial and SCN clocks were monitored using PER2-driven bioluminescence rhythms at the tissue and single-cell levels. We found that polarization of resting microglia to a pro-inflammatory (M1) or anti-inflammatory (M2) state significantly altered the period and amplitude of their molecular circadian clock; importantly, the parameters changed plastically with the repolarization of microglia. This effect was reflected in specific modulations of the expression profiles of individual clock genes in the polarized microglia. Depletion of microglia significantly reduced the amplitude of the SCN clock, and co-cultivation of the SCN explants with M2-polarized microglia specifically improved the amplitude of the SCN clock. These results demonstrate that the presence of M2-polarized microglia has beneficial effects on SCN clock function. Our results provide new insight into the mutual interaction between immune and circadian systems in the brain.
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Data Availability
Python scripts used for image adjustment of bioluminescence microscopy recordings and bioluminescence data analyses are available at: https://github.com/clockgene. The datasets generated during and/or analyzed during the current study are available from the corresponding author on request.
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Acknowledgements
This work was supported by the Grant Agency of Charles University (project GA UK No. 514219), OPPK BrainView CZ.2.16/3.1.00/21544, and Research Project RV0 67985823. The authors thank Eva Tlusta for technical assistance. We would also like to thank Chun-Xia Yi and Irina Milanova (Amsterdam UMC, Amsterdam, Netherlands) for their support and methodology sharing.
Funding
This work was supported by the Grant Agency of Charles University (project GA UK No. 514219), OPPK BrainView CZ.2.16/3.1.00/21544, and Research Project RV0 67985823.
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All authors contributed to the study execution. PH and AS contributed to study conception and design. PH and KS contributed to material preparation, data collection, and analysis. PH contributed to the first draft of the manuscript. AS reviewed and commented on the following versions of the manuscript. All authors read and approved the final manuscript.
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The Animal Care and Use Committee of the Institute of Physiology, in agreement with the Animal Protection Law of the Czech Republic as well as European Community Council directives 86/609/EEC, approved all experiments.
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Supplementary file1 (AVI 1392 KB) Online Resource 1 Bright-field video recording of M0-polarized microglia culture recorded for 96 hours. Snapshots were acquired every 3 hours.
Supplementary file2 (AVI 1210 KB) Online Resource 2 Bright-field video recording of M1-polarized microglia culture recorded for 96 hours. Snapshots were acquired every 3 hours.
Supplementary file3 (AVI 1516 KB) Online Resource 3 Bright-field video recording of M2-polarized microglia culture recorded for 96 hours. Snapshots were acquired every 3 hours.
Supplementary file4 (AVI 4536 KB) Online Resource 4 Bioluminescence video recording of M0-polarized microglia culture recorded for 96 hours. Snapshots were acquired every 3 hours.
Supplementary file5 (AVI 4883 KB) Online Resource 5 Bioluminescence video recording of M1-polarized microglia culture recorded for 96 hours. Snapshots were acquired every 3 hours.
Supplementary file6 (AVI 5009 KB) Online Resource 6 Bioluminescence video recording of M2-polarized microglia culture recorded for 96 hours. Snapshots were acquired every 3 hours.
10571_2022_1252_MOESM7_ESM.pdf
Supplementary file7 (PDF 756 KB) Online Resource 7 Supplemental Figure S1 summarizing obtained results regarding the effect of PPARγ activation/inhibition on PER2-driven bioluminescence rhythms in differently polarized microglia.
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Honzlová, P., Semenovykh, K. & Sumová, A. The Circadian Clock of Polarized Microglia and Its Interaction with Mouse Brain Oscillators. Cell Mol Neurobiol 43, 1319–1333 (2023). https://doi.org/10.1007/s10571-022-01252-1
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DOI: https://doi.org/10.1007/s10571-022-01252-1