Psychotropic drug-induced gene expression alterations in mouse striatum II

To identify the molecular mechanisms that may initiate therapeutic effects, whole-genome expression profiling (Illumina Mouse WG-6 microarrays) of drug-induced alterations in the mouse brain was undertaken, with a focus on the time-course (1, 2, 4 and 8h) of gene expression changes produced by eighteen major psychotropic drugs: antidepressants, antipsychotics, anxiolytics, psychostimulants and opioids. The resulting database is freely accessible at www.genes2mind.org. Bioinformatics approaches led to the identification of three main drug-responsive genomic networks and indicated neurobiological pathways that mediate the alterations in transcription. Each tested psychotropic drug was characterized by a unique gene network expression profile related to its neuropharmacological properties. Functional links that connect expression of the networks to the development of neuronal adaptations (MAPK signaling pathway), control of brain metabolism (adipocytokine pathway), and organization of cell projections (mTOR pathway) were found. The additional data-sets are available at GEOX1 and GEOX2. The microarray experiment was performed to analyze time-course of drug-induced transcriptional response in C57BL/6J mouse striatum. Three antidepressants (bupropion 20 mg/kg, tranylcypromine 20 mg/kg, mianserin 20 mg/kg, i.p.), three anxiolytics (diazepam 5 mg/kg, buspirone 10 mg/kg, hydroxyzine 10 mg/kg, i.p.), and three antipsychotics (clozapine 3 mg/kg, risperidone 0.5 mg/kg, haloperidol 1 mg/kg) were selected for the comparison. Drug doses were previously reported as effective in mice and further tested in our laboratory. To analyze dynamics of early, intermediate and relatively late changes of mRNA abundance the experiment was performed in four time points (1, 2, 4 and 8h after drug administration). To exclude influence of drug injection and circadian rhythm on gene expression profile, control groups of saline or tween (1% Tween 80) treated and naïve animals were prepared for each time point. Design of the experiment assumed pooling of two animals per each array and using of three independent arrays per group. To provide appropriate balance in the whole dataset groups were equally divided between the array hybridization batches.

为探明可介导治疗效应的分子机制，本研究针对18种主要精神药物——包括抗抑郁药、抗精神病药、抗焦虑药、精神兴奋剂与阿片类药物——诱导的小鼠脑部基因表达变化，开展全基因组表达谱分析（采用Illumina Mouse WG-6微阵列），重点考察药物处理后1、2、4、8小时四个时间点的基因表达动态变化过程。
所得研究数据库可于www.genes2mind.org免费获取。
通过生物信息学分析，本研究鉴定出3个主要的药物响应基因组网络，并明确了介导转录改变的神经生物学通路。
每种受试精神药物均具有与其神经药理学特性相匹配的独特基因网络表达谱。
研究还发现了三类功能关联：分别连接网络表达与神经元适应形成（丝裂原活化蛋白激酶（MAPK）信号通路）、脑代谢调控（脂肪细胞因子通路）以及细胞突起组织构建（雷帕霉素靶蛋白（mTOR）通路）。
额外数据集可于GEOX1与GEOX2获取。
本次微阵列实验旨在分析C57BL/6J小鼠纹状体中药物诱导的转录应答时间进程。
本次对比共选取3种抗抑郁药（安非他酮20 mg/kg、反苯环丙胺20 mg/kg、米安色林20 mg/kg，腹腔注射）、3种抗焦虑药（地西泮5 mg/kg、丁螺环酮10 mg/kg、羟嗪10 mg/kg，腹腔注射）及3种抗精神病药（氯氮平3 mg/kg、利培酮0.5 mg/kg、氟哌啶醇1 mg/kg）。
所用药物剂量已在既往小鼠实验中被证实有效，并在本实验室完成了预验证。
为分析mRNA丰度的早期、中期及相对晚期动态变化，实验设置了给药后1、2、4、8小时四个时间点。
为排除药物注射与昼夜节律对基因表达谱的干扰，本研究为每个时间点均设置了生理盐水处理组、1%吐温80处理组及空白对照组动物。
实验设计采用每组2只动物的混合样本进行单张微阵列检测，每组设置3张独立重复微阵列。
为保证全数据集的平衡性，所有实验组均按微阵列杂交批次进行平均分配。