ハイスループット・トランスクリプトミクスを通じてのニューロン構造可塑性規制の分析

在2014财年，我们已经完成了这个项目所需的主要生物信息学分析，使用了使用CAGE实验方案从不同发育阶段的多种果蝇细胞收集的转录组数据。我们决定将重点放在：1)IV类DA神经元的发育过程，以及2)IV类与I-III类细胞类型的规范。我们进行了以下分析：1.根据表达的启动子的序列组成和形状(无论它们是尖锐的峰还是广泛分布的)以及它们是如何差异利用的，对表达的启动子进行鉴定。果蝇特异性转录因子结合位点基序的收集和聚类，并预测它们在果蝇基因组中的位置。由于缺乏现有的果蝇特异性结合基序，我们根据它们的相似性收集和合并基序。基于收集的基序集和可用的芯片序列和芯片数据对差异表达启动子的调控分析。我们已经列出了一系列候选的监管因素，并正在进行实验验证。对分析样本的重复转录组进行分析。我们发现，某些重复序列根据发育阶段或细胞类型的不同而表达不同，我们正在试图确定这些差异背后的调控机制。

For the FY2014, we have completed the main bioinformatics analyses required for this project, using the collected transcriptome data from multiple Drosophila cell types at different stages of development using the CAGE experimental protocol. We decided to focus on: 1) Class IV da neuron development time course, and 2) specification of Class IV vs. the Class I-III cell types. We performed the following analyses:1. Characterization of the expressed promoters based on their sequence composition and shapes (whether they are sharply peaked or broadly spread), and how they are differentially utilized.2. Collecting and clustering of Drosophila-specific transcription factor binding site motifs and predicting their positions on the Drosophila genome. Due to the lack of available Drosophila-specific binding motifs, we collected and merged motifs based on their similarity.3. Regulatory analysis of the differentially expressed promoters based on the collected motif set and available ChIP-seq and ChIP-chip data. We have come up with a list of candidate regulatory factors and are in the process of validating them experimentally.4. Analysis of the repeat transcriptome for the analyzed samples. We found that certain repeats are differentially expressed according to developmental stages or cell types, and we are trying to determine what regulatory controls are behind these differences.