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Functional mapping of the lateral lemniscus and the superior olivary complex as part of the auditory pathway was accomplished for the first time in mice in vivo using manganese-enhanced MRI (2.35T, 3D FLASH, 117 μm isotropic resolution). These and other auditory centers in the brainstem presented with pronounced signal enhancements after systemic administration of manganese chloride when animals were exposed to acoustic stimuli for 48 hr, but not when kept in a quiet environment. The results indicate an activation-dependent accumulation of manganese in the neural circuit composed of the cochlear nucleus, the superior olivary complex, the lateral lemniscus, and the inferior colliculus. The marked enhancement of the lateral lemniscus suggests that the stimulus-related accumulation of manganese reflects not only a regional uptake from extracellular fluid but also a concurrent delivery by axonal transport within the auditory system. Magn Reson Med 60:210–212, 2008. © 2008 Wiley-Liss, Inc.