Wada Kazuhiro, Howard Jason T, McConnell Patrick, Whitney Osceola, Lints Thierry, Rivas Miriam V, Horita Haruhito, Patterson Michael A, White Stephanie A, Scharff Constance, Haesler Sebastian, Zhao Shengli, Sakaguchi Hironobu, Hagiwara Masatoshi, Shiraki Toshiyuki, Hirozane-Kishikawa Tomoko, Skene Pate, Hayashizaki Yoshihide, Carninci Piero, Jarvis Erich D
Department of Neurobiology and Duke Bioinformatics Shared Resource, Duke University Medical Center, Durham, NC 27710, USA.
Proc Natl Acad Sci U S A. 2006 Oct 10;103(41):15212-7. doi: 10.1073/pnas.0607098103. Epub 2006 Oct 3.
Songbirds have one of the most accessible neural systems for the study of brain mechanisms of behavior. However, neuroethological studies in songbirds have been limited by the lack of high-throughput molecular resources and gene-manipulation tools. To overcome these limitations, we constructed 21 regular, normalized, and subtracted full-length cDNA libraries from brains of zebra finches in 57 developmental and behavioral conditions in an attempt to clone as much of the brain transcriptome as possible. From these libraries, approximately 14,000 transcripts were isolated, representing an estimated 4,738 genes. With the cDNAs, we created a hierarchically organized transcriptome database and a large-scale songbird brain cDNA microarray. We used the arrays to reveal a set of 33 genes that are regulated in forebrain vocal nuclei by singing behavior. These genes clustered into four anatomical and six temporal expression patterns. Their functions spanned a large range of cellular and molecular categories, from signal transduction, trafficking, and structural, to synaptically released molecules. With the full-length cDNAs and a lentiviral vector system, we were able to overexpress, in vocal nuclei, proteins of representative singing-regulated genes in the absence of singing. This publicly accessible resource http://songbirdtranscriptome.net can now be used to study molecular neuroethological mechanisms of behavior.
鸣禽拥有用于研究行为脑机制的最为便捷的神经系统之一。然而，鸣禽的神经行为学研究一直受到高通量分子资源和基因操作工具匮乏的限制。为克服这些限制，我们构建了21个来自斑胸草雀大脑的常规、标准化和扣除全长cDNA文库，这些文库涵盖了57种发育和行为条件，旨在尽可能多地克隆大脑转录组。从这些文库中分离出了约14,000个转录本，估计代表4,738个基因。利用这些cDNA，我们创建了一个层次组织的转录组数据库和一个大规模的鸣禽大脑cDNA微阵列。我们使用这些微阵列揭示了一组33个受鸣叫行为调控的在前脑发声核中表达的基因。这些基因聚集成四种解剖学和六种时间表达模式。它们的功能涵盖了从信号转导、运输和结构到突触释放分子等广泛的细胞和分子类别。利用全长cDNA和慢病毒载体系统，我们能够在不鸣叫的情况下在前脑发声核中过代表鸣叫调控基因的蛋白质。这个可公开访问的资源http://songbirdtranscriptome.net现在可用于研究行为的分子神经行为学机制。
