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Increased death of granule neurons in the internal granule cell layer of rev-erbAα mutant mice. (A) Quantification of TUNELpositive cells in the IGL of wild-type and mutant mice. *P<0.05, student t-test. (B) Neuron-specific enolase (NSE) and TUNEL double staining of a (−/−) IGL section. A dying neuron (TUNEL and NSE-positive) is indicated by an arrow. Asterisks denote unstained nuclei of NSE-positive neurons. (C,D) TUNEL staining of cerebellar sections from wild-type (C) and mutant (D) P10 mice. egl, external granule cell layer; ml, molecular layer; pcl, Purkinje cell layer; igl, internal granule cell layer. Bars, 30 µm (B); 125 µm (C,D).
Source publication
The rev-erbA(alpha) gene, belonging to the steroid receptor superfamily of transcription factors, is highly conserved during evolution but little is known so far about its functions in development or in adult physiology. Here, we describe genetically altered mice lacking the rev-erbA(alpha) gene. These animals do not show any obvious phenotype in e...
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... or that had reached the IGL survived in the rev-erbAα −/− mouse. Although no difference could be found between wild-type and mutant animals in the molecular layer, clear differences were observed in the IGL. In normal mice, naturally occurring cell death was higher at P10 and then progressively declined to reach undetectable levels in the adult (Fig. 7A). Interestingly, in the −/− mice, we found an increased number of TUNEL-positive cells in the IGL at P10, P13, P16 and P19 (246,385,181 and 157% of control, respectively) but not in the adult mice (Fig. 7A, compare also C,D). Double stainings performed with antibodies against either the neuron specific enolase (NSE) or the glial ...
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... In normal mice, naturally occurring cell death was higher at P10 and then progressively declined to reach undetectable levels in the adult (Fig. 7A). Interestingly, in the −/− mice, we found an increased number of TUNEL-positive cells in the IGL at P10, P13, P16 and P19 (246,385,181 and 157% of control, respectively) but not in the adult mice (Fig. 7A, compare also C,D). Double stainings performed with antibodies against either the neuron specific enolase (NSE) or the glial fibrillary acidic protein (GFAP) showed that the TUNEL-positive cells in the IGL of wild-type and −/− mice were neurons and not astrocytes (Fig. 7B). Thus our results indicate that the deletion of the rev-erbAα ...
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... (246,385,181 and 157% of control, respectively) but not in the adult mice (Fig. 7A, compare also C,D). Double stainings performed with antibodies against either the neuron specific enolase (NSE) or the glial fibrillary acidic protein (GFAP) showed that the TUNEL-positive cells in the IGL of wild-type and −/− mice were neurons and not astrocytes (Fig. 7B). Thus our results indicate that the deletion of the rev-erbAα gene affects the survival of IGL neurons during postnatal ...
Citations
... Interestingly, the downstream gene targets of E-box include various fibrotic markers such as α-smooth muscle actin (αSMA) and vimentin (VIM) 16 . Moreover, the removal of REV-ERBα has been associated with increased risks of lung inflammation and premature senescence, which has been confirmed by our and others' previous studies [17][18][19] . ...
Molecular clock REV-ERBα is central to regulating lung injuries, and decreased REV-ERBα abundance mediates sensitivity to pro-fibrotic insults and exacerbates fibrotic progression. In this study, we determine the role of REV-ERBα in fibrogenesis induced by bleomycin and Influenza A virus (IAV). Bleomycin exposure decreases the abundance of REV-ERBα, and mice dosed with bleomycin at night display exacerbated lung fibrogenesis. Rev-erbα agonist (SR9009) treatment prevents bleomycin induced collagen overexpression in mice. Rev-erbα global heterozygous (Rev-erbα Het) mice infected with IAV showed augmented levels of collagens and lysyl oxidases compared with WT-infected mice. Furthermore, Rev-erbα agonist (GSK4112) prevents collagen and lysyl oxidase overexpression induced by TGFβ in human lung fibroblasts, whereas the Rev-erbα antagonist exacerbates it. Overall, these results indicate that loss of REV-ERBα exacerbates the fibrotic responses by promoting collagen and lysyl oxidase expression, whereas Rev-erbα agonist prevents it. This study provides the potential of Rev-erbα agonists in the treatment of pulmonary fibrosis.
... Besides, FGF21 was regulated indirectly by the nuclear receptors RER-ERBα, and the circadian protein PER2 modulated the repressive function of RER-ERBα, affecting the expression of FGF21 (Chavan et al. 2017). RER-ERBα is central to regulating complex interactions between the circadian clock and metabolism (Chomez et al. 2000). Otherwise, the FGF21 promoter in liver may be an intermediary between the clock components, metabolism, and physiological activities (Chavan et al. 2017). ...
Deoxynivalenol (DON) is widely emerging in various grain crops, milk, and wine products, which can trigger different toxic effects on humans and animals by inhalation or ingestion. It also imposes a considerable financial loss on the agriculture and food industry each year. Previous studies have reported acute and chronic toxicity of DON in liver, and liver is not only the main detoxification organ for DON but also the circadian clock oscillator directly or indirectly regulates critical physiologically hepatic functions under different physiological and pathological conditions. However, researches on the association of circadian rhythm in DON-induced liver damage are limited. In the present study, mice were divided into four groups (CON, DON, Bmal1OE, and Bmal1OE + DON) and AAV8 was used to activate (Bmal1) expression in liver. Then mice were gavaged with 5 mg/kg bw/day DON or saline at different time points (ZT24 = 0, 4, 8, 12, 16, and 20 h) in 1 day and were sacrificed 30 min after oral gavage. The inflammatory cytokines, signal transducers, and activators of transcription Janus kinase/signal transducers and activator of transcription 3 (JAKs/STAT3) pathway and bile acids levels were detected by enzyme-linked immunosorbent assay (ELISA), western blotting, and target metabolomics, respectively. The DON group showed significantly elevated interleukin-1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) levels (P < 0.05 for both) and impaired liver function with rhythm disturbances compared to the CON and Bmal1OE groups. At the molecular level, expressions of some circadian clock proteins were significantly downregulated (P < 0.05 for both) and JAKs/STAT3 pathway was activated during DON exposure, accompanied by indicated circadian rhythm disturbance and inflammatory damage. Importantly, Bmal1 overexpression attenuated DON-induced liver damage, while related hepatic bile acids such as cholic acid (CA) showed a decreasing trend in the DON group compared with the CON group. Our study demonstrates a novel finding that Bmal1 plays a critical role in attenuating liver damage by inhibiting inflammatory levels and maintaining bile acids levels under the DON condition. Therefore, Bmal1 may also be a potential molecular target for reducing the hepatotoxic effects of DON in future studies.
... 104 With respect to lipid metabolism, Rev-erba À/À mice showed increased very low-density lipoprotein (VLDL) and triglyceride levels, consistent with the observed upregulation of apolipoprotein C-III (Apoc3), a critical regulator in triglyceride metabolism. 105,106 Depletion of both Rev-erbs in the liver synergistically de-repressed several metabolic genes as well as genes that control the positive limb of the molecular clock. 107 Consistent with these genetic results, administration of the REV-ERB agonist SR9009 decreased cholesterol levels in the plasma in both wild-type and lowdensity lipoprotein receptor (Ldlr) null mice through downregulating cholesterol biosynthesis gene expression. ...
The circadian clock is a fundamental biological mechanism that orchestrates essential cellular and physiological processes to optimize fitness and health. The basic functional unit is the cell-autonomous oscillator, consisting of intersecting negative feedback loops. Whereas the core loop is primarily responsible for rhythm generation, auxiliary loops, most notably the secondary or stabilization loop, play pivotal roles to confer temporal precision and molecular robustness. The stabilization loop contains opposing nuclear receptor subfamilies REV-ERBs and retinoic acid receptor-related orphan receptors (RORs), competing to modulate rhythmic expression of the basic helix-loop-helix ARNT like 1 ( Bmal1 ) genes in the core loop as well as other clock-controlled genes. Therefore, REV-ERBs and RORs are strategically located to interface the oscillator and the global transcriptomic network, promoting cellular homeostasis and physiological fitness throughout lifespan. Disruption of REV-ERB and ROR functions has been linked with diseases and aging, and pharmacological manipulation of these factors has shown promise in various mouse disease models. Nobiletin is a natural compound that directly binds to and activates RORα/γ, modulating circadian rhythms, and shows robust in vivo efficacies to combat clock-associated pathophysiologies and age-related decline. Results from several studies demonstrate an inverse relation between nobiletin efficacy and clock functional state, where nobiletin elicits little effect in young and healthy mice with growing efficacy as the clock is perturbed by environmental and genetic challenges. This mode of action is consistent with the function of the stabilization loop to promote circadian and physiological resilience. Future studies should further investigate the function and mechanism of REV-ERBs and RORs, and test strategies targeting these factors against disease and aging.
... 102 With respect to lipid metabolism, Rev-erbaÀ/À mice showed increased very low-density lipoprotein (VLDL) and triglyceride levels, consistent with the observed upregulation of apolipoprotein C-III (Apoc3), a critical regulator in triglyceride metabolism. 103,104 Depletion of both Rev-erbs in the liver synergistically de-repressed several metabolic genes as well as genes that control the positive limb of the molecular clock. 105 Consistent with these genetic results, administration of the REV-ERB agonist SR9009 decreased cholesterol levels in the plasma in both wild-type and lowdensity lipoprotein receptor (Ldlr) null mice through downregulating cholesterol biosynthesis gene expression. ...
The circadian clock is a fundamental biological mechanism that orchestrates essential cellular and physiological processes to optimize fitness and health. The basic functional unit is the cell-autonomous oscillator, consisting of intersecting negative feedback loops. Whereas the core loop is primarily responsible for rhythm generation, auxiliary loops, most notably the secondary or stabilization loop, play pivotal roles to confer temporal precision and molecular robustness. The stabilization loop contains opposing nuclear receptor subfamilies REV-ERBs and retinoic acid receptor-related orphan receptors (RORs), competing to modulate rhythmic expression of the basic helix-loop-helix ARNT like 1 ( Bmal1 ) genes in the core loop as well as other clock-controlled genes. Therefore, REV-ERBs and RORs are strategically located to interface the oscillator and the global transcriptomic network, promoting cellular homeostasis and physiological fitness throughout lifespan. Disruption of REV-ERB and ROR functions has been linked with diseases and aging, and pharmacological manipulation of these factors has shown promise in various mouse disease models. Nobiletin is a natural compound that directly binds to and activates RORα/γ, modulating circadian rhythms, and shows robust in vivo efficacies to combat clock-associated pathophysiologies and age-related decline. Results from several studies demonstrate an inverse relation between nobiletin efficacy and clock functional state, where nobiletin elicits little effect in young and healthy mice with growing efficacy as the clock is perturbed by environmental and genetic challenges. This mode of action is consistent with the function of the stabilization loop to promote circadian and physiological resilience. Future studies should further investigate the function and mechanism of REV-ERBs and RORs, and test strategies targeting these factors against disease and aging.
... Some studies have found that activation of REV-ERBα promoted apoptosis in gastric cancer cells and in palmitate-induced preadipocytes Ivyspring International Publisher [15,16]. On the contrary, deletion of the REV-ERBα gene also increase apoptosis of neurons in internal granule cell layer during postnatal cerebellar development [17]. Therefore, REV-ERBα might be a profound regulator of apoptosis, but whether it also regulate Fas-induced apoptosis remains unknown. ...
... In addition, treatment of GSK4112 inhibited the activation of caspase-3 in SH-SH5Y cells [33]. Previous studies have shown that mice lacking REV-ERBα gene enhanced apoptosis of neurons in the internal granule cell layer during postnatal cerebellar development [17]. These data indicate that REV-ERBα plays an important effect in suppressing apoptosis. ...
Fas-induced apoptosis is a central mechanism of hepatocyte damage during acute and chronic hepatic disorders. Increasing evidence suggests that circadian clock plays critical roles in the regulation of cell fates. In the present study, the potential significance of REV-ERBα, a core ingredient of circadian clock, in Fas-induced acute liver injury has been investigated. The anti-Fas antibody Jo2 was injected intraperitoneally in mice to induce acute liver injury and the REV-ERBα agonist GSK4112 was administered. The results indicated that treatment of GSK4112 decreased the level of plasma ALT and AST, attenuated the liver histological changes, and promoted the survival rate in Jo2-insulted mice. Treatment with GSK4112 also downregulated the activities of caspase-3 and caspase-8, suppressed hepatocyte apoptosis. In addition, treatment with GSK4112 decreased the level of Fas and enhanced the phosphorylation of Akt. In conclusion, treatment with GSK4112 alleviated Fas-induced apoptotic liver damage in mice, suggesting that REV-ERBα agonist might have potential value in pharmacological intervention of Fas-associated liver injury.
... NR1D1 is a circadian clock gene and might interact with the gonadotropin-releasing hormone signalling pathway 48 . Knockout of this gene in mice reduces fertility 49 . TGIF1 is a repressor and reversibly ZFY TGIF2LY PCDH11Y TSPY2 AMELY TBL1Y PRKY TSPY4 TSPY8 TSPY3 TSPY1 TSPY10 TSPY9P USP9Y DDX3Y UTY TMSB4Y VCY VCY1B NLGN4Y CDY2B CDY2A HSFY1 HSFY2 AC007244.1 ...
The accurate and complete assembly of both haplotype sequences of a diploid organism is essential to understanding the role of variation in genome functions, phenotypes, and diseases1. Here, using a trio-binning approach, we present a high-quality, diploid reference genome, with both haplotypes assembled independently at the chromosome level, for the common marmoset (Callithrix jacchus), an important primate model system widely used in biomedical research2,3. The full heterozygosity spectrum between the two haplotypes involves 1.36% of the genome, much higher than the 0.13% indicated by the standard single nucleotide heterozygosity estimation alone. The de novo mutation rate is 0.43 × 10-8 per site per generation, where the paternal inherited genome acquired twice as many mutations as the maternal. Our diploid assembly enabled us to discover a recent expansion of the sex differentiated region and unique evolutionary changes in the marmoset Y chromosome. Additionally, we identified many genes with signatures of positive selection that might have contributed to the evolution of Callithrix biological features. Brain related genes were highly conserved between marmosets and humans, though several genes experienced lineage-specific copy number variations or diversifying selection, providing important implications for the application of marmosets as a model system.
... GSK3B is involved in multiple signaling pathways controlling cellular metabolism, differentiation, death, and survival 40 . The NR1D1 gene, also known as Rev-ErbAα, encodes a transcription factor that is a member of the nuclear receptor subfamily 1; deletion of this gene affects the survival of neurons during postnatal development 41 . We found that in vitro lithium enhanced the expression of BCL2 and GSK3B specifically in LCLs from clinical lithium responders (Fig. 3F,G), and not in other groups. ...
Lithium is an effective, well-established treatment for bipolar disorder (BD). However, the mechanisms of its action, and reasons for variations in clinical response, are unclear. We used neural precursor cells (NPCs) and lymphoblastoid cell lines (LCLs), from BD patients characterized for clinical response to lithium (using the “Alda scale” and “NIMH Retrospective Life chart method”), to interrogate cellular phenotypes related to both disease and clinical lithium response. NPCs from two biologically related BD patients who differed in their clinical response to lithium were compared with healthy controls. RNA-Seq and analysis, mitochondrial membrane potential (MMP), cell viability, and cell proliferation parameters were assessed, with and without in vitro lithium. These parameters were also examined in LCLs from 25 BD patients (16 lithium responders and 9 non-responders), and 12 controls. MMP was lower in both NPCs and LCLs from BD; but it was reversed with in vitro lithium only in LCLs, and this was unrelated to clinical lithium response. The higher cell proliferation observed in BD was unaffected by in vitro lithium. Cell death was greater in BD. However, LCLs from clinical lithium responders could be rescued by addition of in vitro lithium. In vitro lithium also enhanced BCL2 and GSK3B expression in these cells. Our findings indicate cellular phenotypes related to the disease (MMP, cell proliferation) in both NPCs and LCLs; and those related to clinical lithium response (cell viability, BCL2/GSK3B expression) in LCLs.
... Moreover, the stimulation of REV-Erbα regulates the tumor necrosis factor (TNF)-or LPS-induced expression of proinflammatory molecules (interleukin-1β (IL-1β), interleukin-6 (IL- 6), and matrix metalloprotease-9 (MMP-9)) in astrocytes or microglia [16][17][18]. In addition, it is worth noting that Rev-Erbα is closely related to neuronal apoptosis and neurodegeneration in the study of Parkinson's disease and cerebellar development [19,20]. However, little is known about the role of Rev-Erbα in TLE. ...
... Several studies have demonstrated that Rev-Erbα and neuronal apoptosis are inextricably linked. For example, some investigators have found a significant increase in apoptosis of internal granule cell layer neurons in the cerebellum of NR1D1 mutant mice [19]. Additionally, dopaminergic neuronal loss in the substantia nigra pars compacta (SNpc) induced by 6-hydroxydopamine is accelerated and intensified in Rev-Erbα knockout mice [20]. ...
Background:
A hallmark of temporal lobe epilepsy (TLE) is brain inflammation accompanied by neuronal demise. Accumulating evidence demonstrates that Rev-Erbα is involved in regulating neuroinflammation and determining the fate of neurons. Therefore, we studied the expression and cellular distribution of Rev-Erbα in the epileptogenic zone of TLE and the effect of treatment with the Rev-Erbα specific agonist SR9009 in the pilocarpine model.
Methods:
The expression pattern of Rev-Erbα was investigated by western blotting, immunohistochemistry, and immunofluorescence labeling in patients with TLE. Next, the effects of SR9009 on neuroinflammation, neuronal apoptosis, and neuronal loss in the mouse hippocampus 7 days after status epilepticus (SE) were assessed by western blotting, immunofluorescence labeling staining, and TUNEL staining.
Results:
The western blotting, immunohistochemistry, and immunofluorescence labeling results revealed that Rev-Erbα was downregulated in the epileptogenic zone of TLE patients and mainly localized in neurons, astrocytes, and presumably microglia. Meanwhile, the expression of Rev-Erbα was decreased in the hippocampus and temporal neocortex of mice treated with pilocarpine in the early post-SE and chronic phases. Interestingly, the expression of Rev-Erbα in the normal hippocampus showed a 24-h rhythm; however, the rhythmicity was disturbed in the early phase after SE, and this disturbance was still present in epileptic animals. Our further findings revealed that treatment with SR9009 inhibited NLRP3 inflammasome activation, inflammatory cytokine (IL-1β, IL-18, IL-6, and TNF-α) production, astrocytosis, microgliosis, and neuronal damage in the hippocampus after SE.
Conclusions:
Taken together, these results suggested that a decrease in Rev-Erbα in the epileptogenic zone may contribute to the process of TLE and that the activation of Rev-Erbα may have anti-inflammatory and neuroprotective effects.
... GSK3B is involved in multiple signaling pathways controlling cellular metabolism, differentiation, death, and survival 40 . The NR1D1 gene, also known as Rev-ErbAα, encodes a transcription factor that is a member of the nuclear receptor subfamily 1; deletion of this gene affects the survival of neurons during postnatal development 41 . We found that in vitro lithium enhanced the expression of BCL2 and GSK3B specifically in LCLs from clinical lithium responders (Fig. 3F,G), and not in other groups. ...
... 30 ), Nr1d1 −/− (ref. 31 ), Arntl fl (ref. 32 ), Nr3c1 fl (ref. ...
Group 3 innate lymphoid cells (ILC3s) are major regulators of inflammation, infection, microbiota composition and metabolism¹. ILC3s and neuronal cells have been shown to interact at discrete mucosal locations to steer mucosal defence2,3. Nevertheless, it is unclear whether neuroimmune circuits operate at an organismal level, integrating extrinsic environmental signals to orchestrate ILC3 responses. Here we show that light-entrained and brain-tuned circadian circuits regulate enteric ILC3s, intestinal homeostasis, gut defence and host lipid metabolism in mice. We found that enteric ILC3s display circadian expression of clock genes and ILC3-related transcription factors. ILC3-autonomous ablation of the circadian regulator Arntl led to disrupted gut ILC3 homeostasis, impaired epithelial reactivity, a deregulated microbiome, increased susceptibility to bowel infection and disrupted lipid metabolism. Loss of ILC3-intrinsic Arntl shaped the gut ‘postcode receptors’ of ILC3s. Strikingly, light–dark cycles, feeding rhythms and microbial cues differentially regulated ILC3 clocks, with light signals being the major entraining cues of ILC3s. Accordingly, surgically or genetically induced deregulation of brain rhythmicity led to disrupted circadian ILC3 oscillations, a deregulated microbiome and altered lipid metabolism. Our work reveals a circadian circuitry that translates environmental light cues into enteric ILC3s, shaping intestinal health, metabolism and organismal homeostasis.
