title: Neural transcriptome reveals molecular mechanisms for temporal control of vocalization across multiple timescales.
abstract: BACKGROUND
  Vocalization is a prominent social behavior among vertebrates, including in the midshipman fish, an established model for elucidating the neural basis of acoustic communication. Courtship vocalizations produced by territorial males are essential for reproductive success, vary over daily and seasonal cycles, and last up to hours per call. Vocalizations rely upon extreme synchrony and millisecond precision in the firing of a homogeneous population of motoneurons, the vocal motor nucleus (VMN). Although studies have identified neural mechanisms driving rapid, precise, and stable neuronal firing over long periods of calling, little is known about underlying genetic/molecular mechanisms.
  
  
  RESULTS
  We used RNA sequencing-based transcriptome analyses to compare patterns of gene expression in VMN to the surrounding hindbrain across three daily and seasonal time points of high and low sound production to identify candidate genes that underlie VMN's intrinsic and network neuronal properties. Results from gene ontology enrichment, enzyme pathway mapping, and gene category-wide expression levels highlighted the importance of cellular respiration in VMN function, consistent with the high energetic demands of sustained vocal behavior. Functionally important candidate genes upregulated in the VMN, including at time points corresponding to high natural vocal activity, encode ion channels and neurotransmitter receptors, hormone receptors and biosynthetic enzymes, neuromodulators, aerobic respiration enzymes, and antioxidants. Quantitative PCR and RNA-seq expression levels for 28 genes were significantly correlated. Many candidate gene products regulate mechanisms of neuronal excitability, including those previously identified in VMN motoneurons, as well as novel ones that remain to be investigated. Supporting evidence from previous studies in midshipman strongly validate the value of transcriptomic analyses for linking genes to neural characters that drive behavior.
  
  
  CONCLUSIONS
  Transcriptome analyses highlighted a suite of molecular mechanisms that regulate vocalization over behaviorally relevant timescales, spanning milliseconds to hours and seasons. To our knowledge, this is the first comprehensive characterization of gene expression in a dedicated vocal motor nucleus. Candidate genes identified here may belong to a conserved genetic toolkit for vocal motoneurons facing similar energetic and neurophysiological demands.
authors.affiliation: Department of Neurobiology and Behavior, Cornell University, 14853, Ithaca, NY, USA. nf82@cornell.edu.
authors.forename: Ni Y
authors.initials: NY
authors.lastname: Feng
authors.affiliation: Department of Neurobiology and Behavior, Cornell University, 14853, Ithaca, NY, USA. danieljfergus@gmail.com.|Current Address: North Carolina Museum of Natural Sciences, Genomics and Microbiology, 27601, Raleigh, NC, USA. danieljfergus@gmail.com.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Fergus
authors.affiliation: Department of Neurobiology and Behavior, Cornell University, 14853, Ithaca, NY, USA. ahb3@cornell.edu.
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Bass
mesh_terms: D000818:Animals; D024541:Batrachoidiformes / Q000033:anatomy & histology / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D008297:Male; D009046:Motor Neurons / Q000502:physiology; D012249:Rhombencephalon / Q000502:physiology*; D012621:Seasons; D017423:Sequence Analysis, RNA / Q000379:methods*; D012919:Social Behavior; D013997:Time Factors; D014828:Vocalization, Animal