Project description:Background: Vocal learning is a rare and complex behavioral trait that serves as a basis for the acquisition of human spoken language. In songbirds, vocal learning and production depend on a set of specialized brain nuclei known as the song system. Methodology/Principal Findings: Using high-throughput functional genomics we have identified, 200 novel molecular markers of adult zebra finch HVC Vocal, a key node of the song system. These markers clearly differentiate HVC from the general pallial region to which HVC belongs, and thus represent molecular specializations of this song nucleus. Bioinformatics analysis reveals that several major neuronal cell functions and specific biochemical pathways are the targets of transcriptional regulation in HVC, including: 1) cell-cell and cell-substrate interactions (e.g., cadherin/catenin-mediated adherens junctions, collagen-mediated focal adhesions, and semaphorin-neuropilin/plexin axon guidance pathways); 2) cell excitability (e.g., potassium channel subfamilies, cholinergic and serotonergic receptors, neuropeptides and neuropeptide receptors); 3) signal transduction (e.g., calcium regulatory proteins, regulators of G-protein-related signaling); 4) cell proliferation/death, migration and differentiation (e.g., TGF-beta/BMP and p53 pathways); and 5) regulation of gene expression (candidate retinoid and steroid targets, modulators of chromatin/nucleolar organization). The overall direction of regulation suggest that processes related to cell stability are enhanced, whereas proliferation, growth and plasticity are largely suppressed in adult HVC, consistent with the observation that song in this songbird species is mostly stable in adulthood. Conclusions/Significance: Our study represents one of the most comprehensive molecular genetic characterizations of a brain nucleus involved in a complex learned behavior in a vertebrate. The data indicate numerous targets for pharmacological and genetic manipulations of the song system, and provide novel insights into mechanisms that might play a role in the regulation of song behavior and/or vocal learning.

Project description:Background: Vocal learning is a rare and complex behavioral trait that serves as a basis for the acquisition of human spoken language. In songbirds, vocal learning and production depend on a set of specialized brain nuclei known as the song system. Methodology/Principal Findings: Using high-throughput functional genomics we have identified, 200 novel molecular markers of adult zebra finch HVC Vocal, a key node of the song system. These markers clearly differentiate HVC from the general pallial region to which HVC belongs, and thus represent molecular specializations of this song nucleus. Bioinformatics analysis reveals that several major neuronal cell functions and specific biochemical pathways are the targets of transcriptional regulation in HVC, including: 1) cell-cell and cell-substrate interactions (e.g., cadherin/catenin-mediated adherens junctions, collagen-mediated focal adhesions, and semaphorin-neuropilin/plexin axon guidance pathways); 2) cell excitability (e.g., potassium channel subfamilies, cholinergic and serotonergic receptors, neuropeptides and neuropeptide receptors); 3) signal transduction (e.g., calcium regulatory proteins, regulators of G-protein-related signaling); 4) cell proliferation/death, migration and differentiation (e.g., TGF-beta/BMP and p53 pathways); and 5) regulation of gene expression (candidate retinoid and steroid targets, modulators of chromatin/nucleolar organization). The overall direction of regulation suggest that processes related to cell stability are enhanced, whereas proliferation, growth and plasticity are largely suppressed in adult HVC, consistent with the observation that song in this songbird species is mostly stable in adulthood. Conclusions/Significance: Our study represents one of the most comprehensive molecular genetic characterizations of a brain nucleus involved in a complex learned behavior in a vertebrate. The data indicate numerous targets for pharmacological and genetic manipulations of the song system, and provide novel insights into mechanisms that might play a role in the regulation of song behavior and/or vocal learning. Comparison of HVC and shelf regions from adult male zebra finches, 6 biological replicates per group. Each sample was hybridized against the SoNG universal reference RNA pool.

Project description:Studies of transcriptional networks in multi-cellular organisms usually focus on isolated cells and typically assume that the discovered gene networks represent those present in connected cells within a complex organ like the brain. However, similar cell types connected in diverse anatomical networks could differentially influence transcriptional networks. Here, we used laser capture microdissection, expression arrays, genome mapping, and computational inference to explore behaviorally regulated gene networks in the brains of awake, behaving songbirds producing a skilled motor behavior, singing. We found that at baseline, in the absence of singing, a large proportion of genes (17%, >3000) are differentially expressed in the different brain regions of the neural circuit that controls singing. These genes predominantly code for cell communication and connectivity proteins, and non-coding RNAs. Remarkably, the act of singing was associated with differential regulation of ~10% of the coding and non-coding genome. However, less than 1% of genes were singing-regulated in most brain regions and these were largely immediate early genes (IEGs), which peaked early, including the inducible transcription factors EGR1 and FOS. The remaining vast majority of behaviorally regulated gene expression was specific to one or a subset of brain regions, which peaked later. Promoters of the baseline, common, and diverse singing regulated gene clusters were enriched for different combinations of post-translationally activated transcription factors, like CREB, SRF, MEF2, MZF, and the IEG transcription factors. The results suggest that diverse cell-to-cell interactions and differential combinatorial binding of a small group of transcription factors may influence regional diversity of gene networks in seemingly similar cell types. Thus, in highly integrated neural circuits of intact behaving animals, transcriptional network diversity appears to be the rule, rather than the exception. Gene expression in Area X, HVC, LMAN, and Ra was measured before singing (0) or after singing for 0.5, 1, 2, 3, 4, 5, 6, and 7hours. Four-Six independent experiments were performed at each of the 9 timepoints.

Dataset Information

Singing female songbirds: Female-specific gene networks in the song area HVC

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