首页> 外文期刊>Neuroscience: An International Journal under the Editorial Direction of IBRO >DOWNSTREAM EFFECTS OF STRIATAL-ENRICHED PROTEIN TYROSINE PHOSPHATASE REDUCTION ON RNA EXPRESSION IN VIVO AND IN VITRO
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DOWNSTREAM EFFECTS OF STRIATAL-ENRICHED PROTEIN TYROSINE PHOSPHATASE REDUCTION ON RNA EXPRESSION IN VIVO AND IN VITRO

机译:纹状体蛋白酪氨酸磷酸酶还原对动物体内外表达RNA的下游效应

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摘要

Striatal-enriched protein tyrosine phosphatase (STEP) is a brain-specific tyrosine phosphatase that has been shown to de-phosphorylate several key neuronal signaling proteins, including kinases (extracellular signal-regulated kinase (ERK1/2), FYN, PYK2) and glutamate receptor subunits (N-methyl-D-aspartate receptor subtype 2B (NR2B), glutamate receptor 2 (GLUR2)). Step knock-out mice have increased phosphorylation of these substrates in the brain, with potential functional consequences in synaptic plasticity and cognitive tasks. It is therefore of interest to identify the molecular pathways and downstream transcriptional targets that are impacted by Step knockdown. In the present study, striatal RNA samples from Step wild-type, knock-out and heterozygous mice were hybridized to Affymetrix microarray chips and evaluated for transcriptional changes between genotypes. Pathway analysis highlighted Erk signaling and multiple pathways related to neurotrophin signaling, neuronal development and synaptic transmission. Potential genes of interest identified by microarray were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) in the cortex and hippocampus, which shared several transcriptional alterations with the striatum. In order to evaluate Step knockdown in an in vitro system, a panel of genes were evaluated using qRT-PCR in rat cortical neurons that were transduced with lentivirus expressing short hairpin RNA against Step or a non-targeting control. Our data suggest that Step has a role in the expression of immediate early genes relevant to synaptic plasticity, in both in vitro and in vivo systems. (C) 2014 IBRO. Published by Elsevier Ltd. All rights reserved.
机译:纹状体富集的酪氨酸磷酸酶(STEP)是一种大脑特异性的酪氨酸磷酸酶,已被证明能使几种关键的神经元信号蛋白磷酸化,包括激酶(细胞外信号调节激酶(ERK1 / 2),FYN,PYK2)和谷氨酸。受体亚基(N-甲基-D-天冬氨酸受体亚型2B(NR2B),谷氨酸受体2(GLUR2))。敲除小鼠逐步增加了大脑中这些底物的磷酸化,在突触可塑性和认知任务中具有潜在的功能性后果。因此,感兴趣的是鉴定受步骤敲低影响的分子途径和下游转录靶标。在本研究中,将来自Step野生型,敲除小鼠和杂合小鼠的纹状体RNA样品与Affymetrix微阵列芯片杂交,并评估了基因型之间的转录变化。途径分析突出了Erk信号传导和与神经营养蛋白信号传导,神经元发育和突触传递有关的多种途径。通过定量实时聚合酶链反应(qRT-PCR)在皮层和海马中确认了通过微阵列鉴定的潜在潜在基因,这些基因与纹状体共有多个转录改变。为了在体外系统中评估Step敲低,使用qRT-PCR在大鼠皮层神经元中评估了一组基因,这些基因被表达了针对Step或非靶向对照的短发夹RNA的慢病毒转导。我们的数据表明,在体外和体内系统中,Step在与突触可塑性相关的立即早期基因的表达中都有作用。 (C)2014年IBRO。由Elsevier Ltd.出版。保留所有权利。

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