The nuclear receptor rev-erbα controls circadian thermogenic plasticity

ABSTRACT Circadian oscillation of body temperature is a basic, evolutionarily conserved feature of mammalian biology1. In addition, homeostatic pathways allow organisms to protect their core

temperatures in response to cold exposure2. However, the mechanism responsible for coordinating daily body temperature rhythm and adaptability to environmental challenges is unknown. Here

we show that the nuclear receptor Rev-erbα (also known as Nr1d1), a powerful transcriptional repressor, links circadian and thermogenic networks through the regulation of brown adipose

tissue (BAT) function. Mice exposed to cold fare considerably better at 05:00 (Zeitgeber time 22) when Rev-erbα is barely expressed than at 17:00 (Zeitgeber time 10) when Rev-erbα is

abundant. Deletion of _Rev-erbα_ markedly improves cold tolerance at 17:00, indicating that overcoming Rev-erbα-dependent repression is a fundamental feature of the thermogenic response to

cold. Physiological induction of uncoupling protein 1 (Ucp1) by cold temperatures is preceded by rapid downregulation of _Rev-erbα_ in BAT. Rev-erbα represses Ucp1 in a

brown-adipose-cell-autonomous manner and BAT Ucp1 levels are high in _Rev-erbα_-null mice, even at thermoneutrality. Genetic loss of _Rev-erbα_ also abolishes normal rhythms of body

temperature and BAT activity. Thus, Rev-erbα acts as a thermogenic focal point required for establishing and maintaining body temperature rhythm in a manner that is adaptable to

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Phenotyping, Physiology, and Metabolism Core (R. Ahima and R. Dhir) of the Penn Diabetes Research Center (NIH P30 DK19525). We also thank the Small Animal Imaging Facility of the Perelman

School of Medicine at the University of Pennsylvania (E. Blankemeyer). This work was supported by NIH grants R01 DK45586 (M.A.L.) and F-32 DK095563 (Z.G.-H.) and the JPB Foundation. A.B. was

funded by the Novo Nordisk STAR postdoctoral program. AUTHOR INFORMATION Author notes * Dan Feng and Matthew J. Emmett: These authors contributed equally to this work. AUTHORS AND

AFFILIATIONS * Division of Endocrinology, Department of Medicine, Department of Genetics, Diabetes, and Metabolism, Perelman School of Medicine at the University of Pennsylvania,

Philadelphia, 19104, Pennsylvania, USA Zachary Gerhart-Hines, Dan Feng, Matthew J. Emmett, Logan J. Everett, Erika R. Briggs, Anne Bugge & Mitchell A. Lazar * The Institute for Diabetes,

Obesity, and Metabolism, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA Zachary Gerhart-Hines, Dan Feng, Matthew J. Emmett, Logan J.

Everett, Erika R. Briggs, Anne Bugge, Patrick Seale & Mitchell A. Lazar * Department of Physiology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, 19104,

Pennsylvania, USA Emanuele Loro & Tejvir S. Khurana * Pennsylvania Muscle Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

Emanuele Loro & Tejvir S. Khurana * Department of Radiology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA Catherine Hou & 

Daniel A. Pryma * Department of Pediatrics, Children’s Hospital of Philadelphia, Philadelphia, 19104, Pennsylvania, USA Christine Ferrara * Department of Cell and Developmental Biology,

Perelman School of Medicine at the University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA Patrick Seale Authors * Zachary Gerhart-Hines View author publications You can also

search for this author inPubMed Google Scholar * Dan Feng View author publications You can also search for this author inPubMed Google Scholar * Matthew J. Emmett View author publications

You can also search for this author inPubMed Google Scholar * Logan J. Everett View author publications You can also search for this author inPubMed Google Scholar * Emanuele Loro View

author publications You can also search for this author inPubMed Google Scholar * Erika R. Briggs View author publications You can also search for this author inPubMed Google Scholar * Anne

Bugge View author publications You can also search for this author inPubMed Google Scholar * Catherine Hou View author publications You can also search for this author inPubMed Google

Scholar * Christine Ferrara View author publications You can also search for this author inPubMed Google Scholar * Patrick Seale View author publications You can also search for this author

inPubMed Google Scholar * Daniel A. Pryma View author publications You can also search for this author inPubMed Google Scholar * Tejvir S. Khurana View author publications You can also

search for this author inPubMed Google Scholar * Mitchell A. Lazar View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS D.F., M.J.E., L.J.E.,

E.R.B., A.B. and C.F. performed key experiments/data analysis and read the manuscript. P.S. provided advice and read the manuscript. E.L. and T.S.K. designed, performed and analysed EMG

studies and read the manuscript. C.H. and D.A.P. designed, performed and analysed 18FDG scans and read the manuscript. Z.G.H. performed many of the experiments, and Z.G.H. and M.A.L.

conceived the project, designed experiments, analysed all results and wrote the manuscript. CORRESPONDING AUTHOR Correspondence to Mitchell A. Lazar. ETHICS DECLARATIONS COMPETING INTERESTS

The authors declare no competing financial interests. EXTENDED DATA FIGURES AND TABLES EXTENDED DATA FIGURE 1 THE BAT CORE CLOCK IS LARGELY UNAFFECTED BY _REV-ERBΑ_ DELETION. A, Rev-erbα

protein levels in BAT of wild-type and _Rev-erbα_ knockout mice (_n_ = 2; each lane of the western blot represents pooled biological duplicates). B, BAT mRNA for indicated genes from

wild-type and _Rev-erbα_ knockout mice collected at the indicated times over a 24-h time course (_n_ = 3). EXTENDED DATA FIGURE 2 REV-ERBΑ CONTROLS COLD AND NORADRENALINE-INDUCED OXIDATIVE

METABOLISM INDEPENDENTLY OF SKELETAL MUSCLE METABOLISM. A, Food intake from cold-challenged _Rev-erbα_ knockout mice and control littermates in Fig. 1f. B, r.m.s. derivation of EMG

measurement from Fig. 1g. C, Oxygen consumption rates of _Rev-erbα_ KO mice and control littermates following noradrenaline administration (1 mg kg−1 s.c.) (_n_ = 6). D, E, r.m.s. derivation

of EMG measurements performed on wild-type and _Rev-erbα_ knockout mice following noradrenaline administration (1 mg kg−1 s.c.) (_n_ = 4). ***_P_ < 0.001 as determined by Student’s

_t_-test. Data are expressed as mean ± s.d. EXTENDED DATA FIGURE 3 _REV-ERBΑ_, BUT NOT _REV-ERBΒ_, IS DECREASED IN A COLD-DEPENDENT MANNER. A–C, _Rev-erbβ_ (A), _Bmal1_ (B) and _Pgc1a_ (C)

mRNA levels in BAT during a cold-exposure time course (_n_ = 3 for mRNA). D, BAT gene expression following moderate (20 °C) or acute (4 °C) cold challenges (_n_ = 3). E, BAT protein levels

after 3 h noradrenaline administration (1 mg kg−1 i.p.) or cold exposure (_n_ = 3). *_P_ < 0.05, **_P_ < 0.01, ***_P_ < 0.001 as determined by one-way ANOVA with multiple

comparisons and a Tukey’s post-test. Data are expressed as mean ± s.d. EXTENDED DATA FIGURE 4 REV-ERBΑ NEGATIVELY REGULATES _UCP1_. A, B, BAT mRNA (A) and protein (B) from wild-type and

_Rev-erbα_ knockout mice exposed to cold for 6 h as described in Fig. 3a, b. C, mRNA levels in preadipocytes isolated from wild-type mice, differentiated in culture and collected at the

indicated times after synchronization by serum shock (_n_ = 4). **_P_ < 0.01, ***_P_ < 0.001 as determined by one-way ANOVA with multiple comparisons and a Tukey’s post-test. Data are

expressed as mean ± s.d. EXTENDED DATA FIGURE 5 REV-ERBΑ CONTROLS CIRCADIAN OSCILLATION OF SURFACE TEMPERATURE AND BAT ACTIVITY. A, Infrared images from the thermographic surface temperature

analysis performed in Fig. 4c. B, Genotypic differences between BAT and core temperatures from wild-type and _Rev-erbα_ knockout mice acclimated to thermoneutrality (_n_ = 6). C, 18FDG

imaging (_n_ = 4) of _Rev-erbα_ knockout mice and wild-type littermates during the light and dark phases. Representative sagittal planes are shown for each group. *_P_ < 0.05, Δcore

temperature versus ΔBAT temperature; †_P_ < 0.05, core temperature versus _Rev-erbα_ knockout core temperature; ‡_P_ < 0.001, wild-type BAT temperature versus _Rev-erbα_ knockout BAT

temperature as determined by Student’s _t_-test. Data are expressed as mean ± s.e.m. EXTENDED DATA FIGURE 6 THE NUCLEAR RECEPTOR REV-ERBΑ CONTROLS CIRCADIAN THERMOGENIC PLASTICITY. Rev-erbα

regulates the circadian rhythm of body temperature through direct suppression of thermogenesis and BAT activity. Cold exposure during the light phase rapidly overrides Rev-erbα-dependent

repression to induce thermogenic programs. POWERPOINT SLIDES POWERPOINT SLIDE FOR FIG. 1 POWERPOINT SLIDE FOR FIG. 2 POWERPOINT SLIDE FOR FIG. 3 POWERPOINT SLIDE FOR FIG. 4 RIGHTS AND

PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Gerhart-Hines, Z., Feng, D., Emmett, M. _et al._ The nuclear receptor Rev-erbα controls circadian thermogenic

plasticity. _Nature_ 503, 410–413 (2013). https://doi.org/10.1038/nature12642 Download citation * Received: 15 March 2013 * Accepted: 09 September 2013 * Published: 27 October 2013 * Issue

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