Project description:Songbirds possess 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 this limitation, here we generated a resource of full-length cDNAs for gene expression analyses and gene manipulation in songbirds. We constructed 21 regular, normalized, and subtracted full-length cDNA libraries from brains of songbirds in 57 behavioral and developmental conditions in an attempt to clone as much of the brain transcriptome as possible. From these libraries ~14,000 transcripts were isolated, representing an estimated 4,738 genes, or ~40% of the genes expressed in the brain. With these cDNAs, we created a novel draft transcriptome database and large scale songbird brain cDNA microarrays. We used the arrays to reveal a set of 33 genes regulated in brain vocal nuclei by singing behavior. These genes clustered into 4 anatomical and 6 temporal brain expression patterns. Their functions spanned broad range of cellular and molecular categories, including signal transduction, trafficking, structural, and synaptically released molecules. With the full-length cDNAs, we over-expressed proteins of representative singing-regulated genes in vocal nuclei in the absence of singing, using a lentiviral vector system. This resource now allows investigators to comprehensively study molecular neuroethological mechanisms of behavior. Keywords: songbird, zebra finch, transcriptome, learned vocalization, immediate early genes, lentivirus
