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Similar to many teleost fish, squirrelfish (family Holocentridae) produce vocalizations by the contraction of muscles that lead to vibration of the swimbladder. We used biotinylated compounds to identify the position and extent of vocal motor neurons in comparison to additional motor neuron groups, namely those of red and white dorsal epaxial muscle and opercular muscle that are located adjacent to or near the sonic muscle. The sonic motor nucleus (SMN) was located in the caudal medulla and rostral spinal cord in a ventrolateral position with dendrites extending dorsally in a dense bundle along the lateral edge of the medulla and axons exiting via ventral occipital nerve roots. Transneuronal transport of biocytin identified premotor neurons within the SMN and in the medially adjacent reticular formation that projected to the contralateral SMN and more rostrally to the octavolateralis efferent nucleus and nucleus …