The Psychiatric Risk Gene NT5C2 Regulates Adenosine Monophosphate-Activated Protein Kinase Signaling and Protein Translation in Human Neural Progenitor Cells
The Psychiatric Risk Gene NT5C2 Regulates Adenosine Monophosphate-Activated Protein Kinase Signaling and Protein Translation in Human Neural Progenitor Cells
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DOI：
10.1016/j.biopsych.2019.03.977
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发表时间：
2019-07-15
影响因子：
10.6
通讯作者：
Srivastava, Deepak P.
中科院分区：
文献类型：
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作者：
Duarte, Rodrigo R. R.;Bechtel, Nathaniel D.;Srivastava, Deepak P.
背景技术背景：50-核苷酸酶，胞质II基因（NT 5C 2，cN-II）与以精神和心理障碍为特征的疾病相关。常见的精神病风险等位基因在NT 5C 2基因座减少该基因在胎儿和成人大脑中的表达，但下游的生物风险机制仍然habituet.METHODS：NT 5C 2蛋白在人背外侧前额叶皮层和皮质的人神经祖细胞（hNPC）的分布测定使用免疫染色，公开可用的表达数据，和逆转录酶定量聚合酶链反应。使用蛋白质印迹法对腺苷一磷酸活化蛋白激酶（AMPK）α（Thr 172）和核糖体蛋白S6（Ser 235/Ser 236）进行磷酸化定量，以推断AMPK信号传导的活化程度和蛋白质翻译速率。使用RNA干扰在hNPC和黑腹果蝇中诱导敲低。使用微阵列进行hNPCs的转录组学分析，并使用攀爬assay.Results：NT 5C 2的表达在神经发育过程中较高，并在成年人大脑皮层神经元富集的苍蝇运动行为进行了评估。hNPCs中的敲低影响AMPK信号传导，AMPK信号传导是参与能量稳态和蛋白质翻译的主要营养传感机制。在敲低的hNPC中观察到与蛋白质翻译有关的转录变化，并且使用逆转录酶定量聚合酶链反应证实了AMPK信号传导和蛋白质翻译相关基因的表达变化。在果蝇中的击倒与剧烈的攀登imperfication.CONCLUSIONS：我们提供了一个广泛的神经生物学特征的精神病风险基因NT 5C 2，描述其以前未知的作用，在神经干细胞中的AMPK信号转导和蛋白质翻译的调节和它与果蝇运动行为。
BACKGROUND: The 50-nucleotidase, cytosolic II gene (NT5C2, cN-II) is associated with disorders characterized by psychiatric and psychomotor disturbances. Common psychiatric risk alleles at the NT5C2 locus reduce expression of this gene in the fetal and adult brain, but downstream biological risk mechanisms remain elusive.METHODS: Distribution of the NT5C2 protein in the human dorsolateral prefrontal cortex and cortical human neural progenitor cells (hNPCs) was determined using immunostaining, publicly available expression data, and reverse transcriptase quantitative polymerase chain reaction. Phosphorylation quantification of adenosine monophosphate-activated protein kinase (AMPK) alpha (Thr172) and ribosomal protein S6 (Ser235/Ser236) was performed using Western blotting to infer the degree of activation of AMPK signaling and the rate of protein translation. Knockdowns were induced in hNPCs and Drosophila melanogaster using RNA interference. Transcriptomic profiling of hNPCs was performed using microarrays, and motility behavior was assessed in flies using the climbing assay.RESULTS: Expression of NT5C2 was higher during neurodevelopment and was neuronally enriched in the adult human cortex. Knockdown in hNPCs affected AMPK signaling, a major nutrient-sensing mechanism involved in energy homeostasis, and protein translation. Transcriptional changes implicated in protein translation were observed in knockdown hNPCs, and expression changes to genes related to AMPK signaling and protein translation were confirmed using reverse transcriptase quantitative polymerase chain reaction. The knockdown in Drosophila was associated with drastic climbing impairment.CONCLUSIONS: We provide an extensive neurobiological characterization of the psychiatric risk gene NT5C2, describing its previously unknown role in the regulation of AMPK signaling and protein translation in neural stem cells and its association with Drosophila melanogaster motility behavior.