Serotonin distribution in the brain of the plainfin midshipman: Substrates for vocal‐acoustic modulation and a reevaluation of the serotonergic system in teleost fishes

Serotonin (5‐HT) is a modulator of neural circuitry underlying motor patterning, homeostatic control, and social behavior. While previous studies have described 5‐HT distribution in various teleosts, serotonergic raphe subgroups in fish are not well defined and therefore remain problematic for cross‐species comparisons. Here we used the plainfin midshipman fish, Porichthys notatus, a well‐studied model for investigating the neural and hormonal mechanisms of vertebrate vocal‐acoustic communication, to redefine raphe subgroups based on both stringent neuroanatomical landmarks as well as quantitative cell measurements. In addition, we comprehensively characterized 5‐HT‐immunoreactive (‐ir) innervation throughout the brain, including well‐delineated vocal and auditory nuclei. We report neuroanatomical heterogeneity in populations of the serotonergic raphe nuclei of the brainstem reticular formation, with three discrete subregions in the superior raphe, an intermediate 5‐HT‐ir cell cluster, and an extensive inferior raphe population. 5‐HT‐ir neurons were also observed within the vocal motor nucleus (VMN), forming putative contacts on those cells. In addition, three major 5‐HT‐ir cell groups were identified in the hypothalamus and one group in the pretectum. Significant 5‐HT‐ir innervation was found in components of the vocal pattern generator and cranial motor nuclei. All vocal midbrain nuclei showed considerable 5‐HT‐ir innervation, as did thalamic and hindbrain auditory and lateral line areas and vocal‐acoustic integration sites in the preoptic area and ventral telencephalon. This comprehensive atlas offers new insights into the organization of 5‐HT nuclei in teleosts and provides neuroanatomical evidence for serotonin as a modulator of vocal‐acoustic circuitry and behavior in midshipman fish, consistent with findings in vocal tetrapods. We comprehensively characterized 5‐HT‐immunoreactive (‐ir) innervation throughout the brain of Porichthys notatus, plainfin midshipman fish, including well‐delineated vocal and auditory nuclei. We report neuroanatomical heterogeneity in populations of the serotonergic raphe nuclei of the brainstem reticular formation, with three discrete subregions in the superior raphe, an intermediate 5‐HT‐ir cell cluster, and an extensive inferior raphe population. This comprehensive atlas offers new insights into the organization of 5‐HT nuclei in teleosts and provides neuroanatomical evidence for serotonin as a modulator of vocal‐acousti