Abstract There is increasing evidence that astrocytes play important roles in immune regulation the brain. Astrocytes express toll‐like receptors (TLR) and build up responses to innate triggers by releasing proinflammatory molecules. We investigate signaling pathways released molecules after astrocyte TLR4 activation. Purified rodent brain cultures were treated with activator bacterial lipopolysaccharide (LPS). Tools used interfere this system include small interference RNA, inhibitory...

The p38 mitogen-activated protein kinase (MAPK) signaling pathway, acting through the downstream MK2, regulates stability of many proinflammatory mRNAs that contain adenosine/uridine-rich elements (AREs). It is thought to do this by modulating expression or activity ARE-binding proteins regulate mRNA turnover. MK2 phosphorylates and mRNA-destabilizing tristetraprolin (TTP) at serines 52 178. Here we show MAPK pathway subcellular localization TTP protein. A inhibitor causes rapid...

The expression of GLUT-1 (erythroid/Hep G2) and GLUT-4 (muscle/fat) glucose transporters was assessed during development in rat heart, skeletal muscle, brown adipose tissue. protein detectable fetal heart by day 21 pregnancy; it increased progressively after birth, attaining levels close to those adults at 15 post natal. In contrast, messenger RNA (mRNA) already present hearts from 17 day-old fetuses. mRNA stayed low early postnatal life tissue only 10 pattern for muscle comparable that...

The Zfp36l1 gene encodes a zinc finger-containing mRNA binding protein implicated in the posttranscriptional control of expression. Mouse embryos homozygous for targeted mutation locus died mid-gestation and exhibited extraembryonic intraembryonic vascular abnormalities heart defects. In developing placenta, there was failure mesoderm to invaginate trophoblast layer. phenotype associated with an elevated expression endothelial growth factor (VEGF)-A embryonic fibroblasts cultured under...

COX-2 (cyclo-oxygenase-2) mRNA is degraded rapidly in resting cells, but stabilized by the mitogen-activated protein kinase p38 signalling pathway response to pro-inflammatory stimuli. A conserved ARE (AU-rich element) of 3´ untranslated region, CR1 (conserved region 1), acts as a potent instability determinant, and mediates stabilization activation. detailed structural functional analysis this element was performed an attempt identify RNA-binding proteins involved regulation stability....

Hypoxia-inducible factor (HIF) is a heterodimeric transcription that activates the cellular response to hypoxia. The HIF1alpha subunit constantly synthesized and degraded under normoxia, but degradation rapidly inhibited when oxygen levels drop. Oxygen-dependent hydroxylation by prolyl-4-hydroxylases (PHD) mediates proteasome degradation. Brain ischemia limits availability not only of also glucose. We hypothesized this circumstance could have modulating effect on HIF. assessed separate...

We have reported a novel functional co-operation among MyoD, myocyte enhancer factor-2 (MEF2), and the thyroid hormone receptor in muscle-specific of rat GLUT4 gene muscle cells. Here, we demonstrate that operates skeletal is fiber-dependent innervation-independent. Under normal conditions, both soleus extensor digitorum longus muscles, activity required integrity MEF2-binding site. Cancellation binding site enhanced its activity, suggesting an inhibitory role. Muscle regeneration muscles...

Glucose constitutes a major fuel for the heart, and high glucose uptake during fetal development is coincident with highest level of expression transporter GLUT-1 life. We have previously reported that repressed perinatally in rat GLUT-4, which shows low stage, becomes main adult. Here, we show perinatal GLUT-4 transporters heart controlled directly at gene transcription. Transient transfection assays −99/−33 fragment <i>GLUT-1</i> sufficient to drive transcriptional activity neonatal...

Short interfering RNA (siRNA) inhibits the synthesis of specific proteins through interference (RNAi). However, siRNA can induce innate immune responses that are mediated by toll-like receptors (TLRs) in cells system. Here, we sought to evaluate whether such glial cells. We examined effects various sequences prepared with lipids (oligofectamine). Lipid-siRNA induced variable degrees silencing-independent nonspecific effects, e.g. increased Stat1 and Cox-2 expression release IL-6 IP-10...

Histone deacetylase SIRT1 represses gene expression through the deacetylation of histones and transcription factors is involved in protective cell response to stress aging. However, upon endoplasmic reticulum (ER) stress, impairs IRE1α branch unfolded protein (UPR) inhibition transcriptional activity XBP-1 deficiency beneficial under these conditions. We hypothesized that may unlock blockade unrelated UPR promoting synthesis chaperones improving stability immature proteins or triggering...

Abstract We have previously reported that innervation-dependent basal contractile activity regulates in an inverse manner the expression of GLUT1 and GLUT4 glucose transporters skeletal muscle. Based on facts muscle innervation decreases denervation increases cAMP levels, we investigated whether might mediate effects innervation/denervation transporter expression. Treatment L6E9 myotubes with 8-bromo-cAMP, forskolin, or monobutyryl-8-bromo-cAMP led to a marked decrease protein levels;...

We have studied the activity of system A transport in skeletal muscle during experimental diabetes. Five days after streptozotocin injection, rats showed a marked hyperglycemia and substantial decrease content GLUT‐4 protein adipose tissue. Under these conditions, basal uptake 2‐(methyl)aminoisobutyric acid (MeAIB), an index activity, was enhanced extensor digitorum longus (EDL) muscles from diabetic compared to controls. Furthermore, insulin‐stimulated MeAIB by incubated EDL soleus markedly...

Glucose consumption is severely depressed in the ischemic core, whereas it maintained or even increased penumbral regions during ischemia. Conversely, glucose utilization reduced early after reperfusion spite that and oxygen are available. Experimental studies suggest hypometabolism might be an predictor of brain infarction. However, relationship between with later development infarction remains to further studied same subjects. Here, was assessed vivo by positron emission tomography (PET)...

Serum is among the agents known to induce hypertrophy of cardiac myocytes, which occurs concomitant with an increase in AP-1-mediated transcription. We have examined if this effect correlates changes relative abundance particular AP-1 heterodimers, as their exact composition under these conditions unknown. Furthermore, we obtained insight on specific role c-Fos from studying induction glucose transporter GLUT1 by serum fibroblasts.We characterised heterodimers expressed neonatal myocytes...

Abstract Brain ischemia causes neuronal cell death by several mechanisms involving necrotic and apoptotic processes. The contributions of each process depend on conditions such as the severity duration ischemia, availability ATP. We examined whether glucose affected development apoptosis after transient this was sensitive to caspase inhibition. Retinoic acid‐differentiated SH‐SY5Y human neuroblastoma cells were subjected oxygen deprivation for 15 h followed various periods reoxygenation in...

Abstract Background Valuable clone collections encoding the complete ORFeomes for some model organisms have been constructed following completion of their genome sequencing projects. These libraries are based on Gateway cloning technology, which facilitates study protein function by simplifying subcloning open reading frames (ORF) into any suitable destination vector. The expression proteins interest as fusions with functional modules is a frequent approach in initial characterization. A...

Gene promoter activity can be studied in vivo by molecular imaging methods using reporter gene technology. Transcription of the and reported genes occurs simultaneously. However, depends on protein translation, stability, cellular fate that may differ among various proteins. A double transgenic mouse strain expressing firefly luciferase (lucF) fluorescent mPlum under transcriptional control thermo-inducible heat-shock (Hspa1b) was generated allowing to follow up proteins different...