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Nucleus ambiguus (NA) consists of the glossopharyngeal (CN X) and vagus (CN IX) nerves. The name ‘ambiguus’ derives from the difficulty of finding it and its variation by species. In the medulla oblongata, NA is located in the reticular formation. NA gives rise to the efferent motor fibers of the vagus nerve (CN X) terminating in the laryngeal and pharyngeal muscles. NA and the dorsal motor nucleus of the vagus (DMNV) innervate distinct populations of cardiac ganglionic principal neurons. NA is located just besides the rostral ventrolateal medulla and the vagus fibers driven from DMNV in transversely sectioned medullary slices.