Generation of a conditional knockout allele for the NFAT5 gene in mice

Küper, Christoph; Beck, Franz‐Xaver; Neuhofer, Wolfgang

doi:10.3389/fphys.2014.00507

Cited by 24 publications

(37 citation statements)

References 28 publications

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“…Using loss-of-function experiments, naïve CD4 + T cells from NFAT5 knockout (NFAT5ko) mice (34) showed a significantly increased T reg induction potential [CD4 + CD25 + FoxP3 + T regs (percentage of CD4 + T cells), 17.1 ± 2.8 versus 30.3 ± 1.2, P < 0.01; Fig. 5, A and B].…”

Section: Resultsmentioning

confidence: 99%

A miRNA181a/NFAT5 axis links impaired T cell tolerance induction with autoimmune type 1 diabetes

et al. 2018

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Molecular checkpoints that trigger the onset of islet autoimmunity or progression to human type 1 diabetes (T1D) are incompletely understood. Using T cells from children at an early stage of islet autoimmunity without clinical T1D, we find that a microRNA181a (miRNA181a)–mediated increase in signal strength of stimulation and costimulation links nuclear factor of activated T cells 5 (NFAT5) with impaired tolerance induction and autoimmune activation. We show that enhancing miRNA181a activity increases NFAT5 expression while inhibiting FOXP3+ regulatory T cell (Treg) induction in vitro. Accordingly, Treg induction is improved using T cells from NFAT5 knockout (NFAT5ko) animals, whereas altering miRNA181a activity does not affect Treg induction in NFAT5ko T cells. Moreover, high costimulatory signals result in phosphoinositide 3-kinase (PI3K)–mediated NFAT5, which interferes with FoxP3+ Treg induction. Blocking miRNA181a or NFAT5 increases Treg induction in murine and humanized models and reduces murine islet autoimmunity in vivo. These findings suggest targeting miRNA181a and/or NFAT5 signaling for the development of innovative personalized medicines to limit islet autoimmunity.

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Section: Resultsmentioning

confidence: 99%

A miRNA181a/NFAT5 axis links impaired T cell tolerance induction with autoimmune type 1 diabetes

et al. 2018

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“…To investigate the relevance of nfat5 for smooth muscle cell responses in vivo, we crossed mice carrying an N5 fl/fl (18) with mice expressing a tamoxifen-inducible CreER T2 -recombinase under the control of the SM-MHC promoter (19). NFAT5 protein was barely detectable in arteries under homeostasis except for the aorta of N5 fl/fl mice.…”

Section: Smc-specific Ablation Of the Nfat5 Gene Has No Impact On Artmentioning

confidence: 99%

“…However, despite the fact that mechanoactivation of VSMCs poses as 1 critical stimulus initiating an extensive adaption of their transcriptome, the relevance of NFAT5 for the outcome of biomechanically induced arterial remodeling processes has not been investigated. To this end, we generated a mouse model to genetically ablate Nfat5 in smooth muscle cells (SMCs) (18,19) for the study of its impact on flow-induced collateral growth and pressureinduced hypertrophy of conduit arteries.…”

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confidence: 99%

“…Primary antibodies usedAll animal studies were performed with permission from the Regional Council Karlsruhe (Permission Number.81/ G-124/14) and conformed to the Guide for the Care and Use of Laboratory Animals (NIH). Tonicity enhancer-binding N5 fl/flmice(18) were crossed with smooth muscle-myosin heavy chain The genetic ablation of NFAT5 in the corresponding offspring was induced in male mice (age 10-12 wk) by application of 1 mg tamoxifen per day intraperitoneally on 5 consecutive days [SMC-specific, tamoxifen-induced Nfat5 knockout (N5 (SM)2/2 )] or Miglyol as…”

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Genetic ablation of NFAT5/TonEBP in smooth muscle cells impairs flow‐ and pressure‐induced arterial remodeling in mice

et al. 2018

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The arterial wall adapts to alterations in blood flow and pressure by remodeling the cellular and extracellular architecture. Biomechanical stress of vascular smooth muscle cells (VSMCs) in the media is thought to precede this process and promote their activation and subsequent proliferation. However, molecular determinants orchestrating the transcriptional phenotype under these conditions have been insufficiently studied. We identified the transcription factor, nuclear factor of activated T cells 5 (NFAT5; or tonicity enhancer‐binding protein) as a crucial regulatory element of mechanical stress responses of VSMCs. Here, the relevance of NFAT5 for arterial growth and thickening is investigated in mice upon inducible smooth muscle cell (SMC)‐specific genetic ablation of Nfat5. In cultured mouse VSMCs, loss of Nfat5 inhibits the expression of gene sets involved in the control of the cell cycle and the interaction with the extracellular matrix and cytoskeletal dynamics. In vivo, SMC‐specific knockout of Nfat5 did not affect the general vascular architecture and blood pressure levels under baseline conditions. However, proliferation of VSMCs and the thickening of the arterial wall were inhibited during both flow‐induced collateral remodeling and hypertension‐mediated arterial hypertrophy. Whereas originally described as a hypertonicity‐responsive transcription factor, these findings identify NFAT5 as a novel molecular determinant of biomechanically induced phenotype changes of VSMCs and wall stress‐induced arterial remodeling processes.—Arnold, C., Feldner, A., Zappe, M., Komljenovic, D., De La Torre, C., Ruzicka, P., Hecker, M., Neuhofer, W., Korff, T. Genetic ablation of NFAT5/TonEBP in smooth muscle cells impairs flow‐ and pressure‐induced arterial remodeling in mice. FASEB J. 33, 3364–3377 (2019). http://www.fasebj.org

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“…The original transcription factor for ORE/TonE was identified as TonE-binding protein (TonEBP)/ORE-binding protein (OREBP), which shows sequence homology to a family of transcription factors called the Nuclear Factor of Activated T cells (NFAT), and therefore it was named NFAT5 (Lopez-Rodríguez et al, 1999; Miyakawa et al, 1999; Ko et al, 2000). NFAT5 has been discovered to be ubiquitously expressed throughout the body and has been shown to be sensitive to hypertonicity in all cell types (Lopez-Rodríguez et al, 1999; Trama et al, 2000; Franchi-Gazzola et al, 2001; Zhang et al, 2003; Maallem et al, 2006; Tsai et al, 2006; Küper, Beck & Neuhofer, 2015). However, NFAT5-dependent expression of aldose reductase, BGT1, and SMIT is especially important for the survival of cells in the kidney, as these cells are exposed to high and varied levels of sodium and urea (Miyakawa et al, 1998; Na et al, 2003; López-Rodríguez et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Osmolyte regulation by TonEBP/NFAT5 during anoxia-recovery and dehydration–rehydration stresses in the freeze-tolerant wood frog (Rana sylvatica)

Al-attar

Zhang

2017

PeerJ

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BackgroundThe wood frog, Rana sylvatica, tolerates freezing as a means of winter survival. Freezing is considered to be an ischemic/anoxic event in which oxygen delivery is significantly impaired. In addition, cellular dehydration occurs during freezing because water is lost to extracellular compartments in order to promote freezing. In order to prevent severe cell shrinkage and cell death, it is important for the wood frog to have adaptive mechanisms for osmoregulation. One important mechanism of cellular osmoregulation occurs through the cellular uptake/production of organic osmolytes like sorbitol, betaine, and myo-inositol. Betaine and myo-inositol are transported by the proteins BGT-1 and SMIT, respectively. Sorbitol on the other hand, is synthesized inside the cell by the enzyme aldose reductase. These three proteins are regulated at the transcriptional level by the transcription factor, NFAT5/TonEBP. Therefore, the objective of this study was to elucidate the role of NFAT5/TonEBP in regulating BGT-1, SMIT, and aldose reductase, during dehydration and anoxia in the wood frog muscle, liver, and kidney tissues.MethodsWood frogs were subjected to 24 h anoxia-4 h recovery and 40% dehydration-full rehydration experiments. Protein levels of NFAT5, BGT-1, SMIT, and aldose reductase were studied using immunoblotting in muscle, liver, and kidney tissues.ResultsImmunoblotting results demonstrated downregulations in NFAT5 protein levels in both liver and kidney tissues during anoxia (decreases by 41% and 44% relative to control for liver and kidney, respectively). Aldose reductase protein levels also decreased in both muscle and kidney tissues during anoxia (by 37% and 30% for muscle and kidney, respectively). On the other hand, BGT-1 levels increased during anoxia in muscle (0.9-fold compared to control) and kidney (1.1-fold). Under 40% dehydration, NFAT5 levels decreased in liver by 53%. Aldose reductase levels also decreased by 42% in dehydrated muscle, and by 35% in dehydrated liver. In contrast, BGT-1 levels increased by 1.4-fold in dehydrated liver. SMIT levels also increased in both dehydrated muscle and liver (both by 0.8-fold).DiscussionOverall, we observed that osmoregulation through an NFAT5-mediated pathway is both tissue- and stress-specific. In both anoxia and dehydration, there appears to be a general reduction in NFAT5 levels resulting in decreased aldose reductase levels, however BGT-1 and SMIT levels still increase in certain tissues. Therefore, the regulation of osmoregulatory genes during dehydration and anoxia occurs beyond the transcriptional level, and it possibly involves RNA processing as well. These novel findings on the osmoregulatory mechanisms utilized by the wood frog advances our knowledge of osmoregulation during anoxia and dehydration. In addition, these findings highlight the importance of using this model to study molecular adaptations during stress.

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Generation of a conditional knockout allele for the NFAT5 gene in mice

Cited by 24 publications

References 28 publications

A miRNA181a/NFAT5 axis links impaired T cell tolerance induction with autoimmune type 1 diabetes

A miRNA181a/NFAT5 axis links impaired T cell tolerance induction with autoimmune type 1 diabetes

Genetic ablation of NFAT5/TonEBP in smooth muscle cells impairs flow‐ and pressure‐induced arterial remodeling in mice

Osmolyte regulation by TonEBP/NFAT5 during anoxia-recovery and dehydration–rehydration stresses in the freeze-tolerant wood frog (Rana sylvatica)

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