Seizure control through genetic and pharmacological manipulation of Pumilio in Drosophila: a key component of neuronal homeostasis.
Seizure control through genetic and pharmacological manipulation of Pumilio in Drosophila: a key component of neuronal homeostasis.
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DOI：
10.1242/dmm.027045
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发表时间：
2017-02-01
影响因子：
4.3
通讯作者：
Baines RA
中科院分区：
文献类型：
--
作者：
Lin WH;Giachello CN;Baines RA
癫痫是一种严重的疾病，大约三分之一的患者对药物治疗没有反应。与新靶点相互作用的下一代药物需要为这些人提供更好的临床结果。为了确定抗癫痫药物(AED)设计的潜在新靶点，我们使用RNA测序来确定果蝇黑腹果蝇两种癫痫发作模型中基因转录的变化。第一个模型比较了野生型(WT)和bangsenseless1(Parabss)之间的基因转录，bangsenseless1是唯一的苍蝇电压门控钠通道(麻痹)中的功能获得突变体。第二个模型比较了WT和WT喂饲致痫前胡椒毒素(PTX)。我们鉴定了743个基因(FDR≤1%)，这些基因的表达水平在两种癫痫模型中都是共同的。在这些基因中，339个持续上调，397个下调。我们发现Pumilio(PUM)在两种癫痫模型中都下调了。PUM是已知的苍蝇和哺乳动物动作电位放电的动态平衡调节因子，通过与电压门控钠通道(NAV)的mRNA转录产物结合并调节其翻译来实现对神经元放电的控制。我们发现，维持PUM在Parabss果蝇中枢神经系统中的表达是有效的抗惊厥作用，而通过RNAi介导的敲除它的减少是促进惊厥的。使用基于细胞的荧光素酶报告筛选，我们筛选了一个重新定位的化学文库，并鉴定了12个足以提高PUM活性的化合物。在这些化合物中，我们专注于阿沃苯宗，它显著挽救了鹦鹉的癫痫行为。阿沃苯宗的作用模式包括增强PUM的表达，并反映了这种自我平衡调节因子降低已识别神经元持续电压门控钠电流(INAP)的能力。这项研究报道了一种抑制癫痫发作的新方法，并强调了神经元动态平衡的机制作为下一代抗癫痫药物的潜在靶点。摘要：下一代抗惊厥化合物可增强神经元内稳态调节剂Pumilio的活性。
Epilepsy is a significant disorder for which approximately one-third of patients do not respond to drug treatments. Next-generation drugs, which interact with novel targets, are required to provide a better clinical outcome for these individuals. To identify potential novel targets for antiepileptic drug (AED) design, we used RNA sequencing to identify changes in gene transcription in two seizure models of the fruit fly Drosophila melanogaster. The first model compared gene transcription between wild type (WT) and bangsenseless1 (parabss), a gain-of-function mutant in the sole fly voltage-gated sodium channel (paralytic). The second model compared WT with WT fed the proconvulsant picrotoxin (PTX). We identified 743 genes (FDR≤1%) with significant altered expression levels that are common to both seizure models. Of these, 339 are consistently upregulated and 397 downregulated. We identify pumilio (pum) to be downregulated in both seizure models. Pum is a known homeostatic regulator of action potential firing in both flies and mammals, achieving control of neuronal firing through binding to, and regulating translation of, the mRNA transcripts of voltage-gated sodium channels (Nav). We show that maintaining expression of pum in the CNS of parabss flies is potently anticonvulsive, whereas its reduction through RNAi-mediated knockdown is proconvulsive. Using a cell-based luciferase reporter screen, we screened a repurposed chemical library and identified 12 compounds sufficient to increase activity of pum. Of these compounds, we focus on avobenzone, which significantly rescues seizure behaviour in parabss flies. The mode of action of avobenzone includes potentiation of pum expression and mirrors the ability of this homeostatic regulator to reduce the persistent voltage-gated Na+ current (INaP) in an identified neuron. This study reports a novel approach to suppress seizure and highlights the mechanisms of neuronal homeostasis as potential targets for next-generation AEDs. Summary: Next-generation anticonvulsant compounds potentiate the activity of the neuronal homeostatic regulator Pumilio.