diff), and clustering coefficient of genes annotated by terms with the highest TS scores were compared to the rest of the module, allowing us to home in on particularly tight-knit, GABA inhibition behaviorally relevant, biological pathways/functions in the singing-related modules (Supplemental Experimental Procedures). For example, 11 genes in the blue module (ARC, CABP1, CNN3, DLG1, DLG2, DLGAP2, FREQ, HOMER1, IFNGR1, NLGN1, and NTRK2) were annotated by the term “GO:0014069∼postsynaptic density” ( Table S4). Probes representing these genes in the blue module had high MM and GS.motifs.X (27 probes
total; mean MM = 0.804, GS.motifs.X = 0.682), and the term “GO:0014069∼postsynaptic density” had an enrichment p value of 0.059. Thus TS for this term = 0.804 × 0.682 × (1 − 0.059) = 0.516 (7th highest Lumacaftor of 402 enriched blue module terms; Table S2. Area X Network Data and Table S4. Functional Annotation of Selected Modules). Compared to the rest of the module, probes for the 11 genes annotated with this term had higher average MM (p = 6.2e-7, Kruskal-Wallis test), GS.motifs.X (p = 6.8e-5), kIN.diff (p = 4.7e-6), and clustering coefficient (p = 5.2e-5). Other top-ranked blue module terms included “GO:0031434∼mitogen-activated protein kinase kinase binding” and “IPR019583:PDZ-associated domain of
NMDA receptors,” as well as others involving actin, cytoskeleton, and tyrosine phosphatase regulation. Genes associated with these synapse-related functions in the blue module were also some of FOXP2′s closest neighbors, i.e., genes with which it had high TO ( Figures 6D–6F, Table S2, Supplemental Experimental Procedures). This may imply a role for FoxP2 in the Etomidate suppression of synaptic plasticity, since blue module genes (whose levels increased with singing in these experiments) in high TO with FOXP2 (which decreased with singing) are good candidates for repressed transcriptional targets. Each of the song modules was enriched for astrocytic markers with developing astrocytes most enriched in the blue module (p = 7.5e-6, Fisher’s
exact test) and mature astrocytes in the orange module (p = 4e-3; Cahoy et al., 2008). This observation is consistent with the recent realization that astrocytes are involved in the regulation of neuronal functions, including behavior (Halassa and Haydon, 2010). We screened the modules for genes associated with Parkinson’s disease (Supplemental Experimental Procedures), since it is a basal ganglia based disorder with a vocal component and found enrichment in the black singing-related module (Figure S6). Another module that was moderately singing-related was also enriched for Parkinson’s disease-associated genes, as well as autism susceptibility genes (purple module, p = 2.7e-4, p = 0.05, respectively, Table S2). The unique presence of the song modules in area X implies that the biological pathways they represent are coregulated in patterns specific to area X during learned vocal-motor behavior.