Double knockin mice show NF-?B trajectories in immune signaling and aging

Author:

Rahman, Shah Md Toufiqur

Aqdas, Mohammad

Martin, Erik W

Tomassoni Ardori,Francesco

Songkiatisak, Preeyaporn

Oh, Kyu-Seon

Uderhardt, Stefan

Yun, Sangwon

Claybourne, Quia C

McDevitt, Ross A

Greco, Valentina

Germain, Ronald N

Tessarollo, Lino

Sung, Myong-Hee
Author Address

Laboratory of Molecular Biology and Immunology, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA., Mouse Cancer Genetics Program, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA., Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA., School of Medicine, Yale University, New Haven, CT 06510, USA., Comparative Medicine Section, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA., Laboratory of Molecular Biology and Immunology, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA. Electronic address: sungm@nih.gov.,

Abstract:

In vitro studies suggest that mapping the spatiotemporal complexity of nuclear factor ?B (NF-?B) signaling is essential to understanding its function. The lack of tools to directly monitor NF-?B proteins in vivo has hindered such efforts. Here, we introduce reporter mice with the endogenous RelA (p65) or c-Rel labeled with distinct fluorescent proteins and a double knockin with both subunits labeled. Overcoming hurdles in simultaneous live-cell imaging of RelA and c-Rel, we show that quantitative features of signaling reflect the identity of activating ligands, differ between primary and immortalized cells, and shift toward c-Rel in microglia from aged brains. RelA:c-Rel heterodimer is unexpectedly depleted in the nuclei of stimulated cells. Trajectories of subunit co-expression in immune lineages reveal a reduction at key cell maturation stages. These results demonstrate the power of these reporters in gaining deeper insights into NF-?B biology, with the spectral complementarity of the labeled NF-?B proteins enabling diverse applications. Published by Elsevier Inc.

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