Preference behaviors are often established during early life, but the underlying neural circuit mechanisms remain unknown. Adapting a unique nesting behavior assay, we confirmed a "critical period" for developing music preference in C57BL/6 mice. Early music exposure between postnatal days 15 and 24 reversed their innate bias for silent shelter, which typically could not be altered in adulthood. Instead, exposing adult mice treated acutely with valproic acid or carrying a targeted deletion of the Nogo receptor (NgR) unmasked a strong plasticity of preference consistent with a reopening of the critical period as seen in other systems. Imaging of cFos expression revealed a prominent neuronal activation in response to the exposed music in the prelimbic and infralimbic medial prefrontal cortex only under conditions of open plasticity. Neither behavioral changes nor selective medial prefrontal cortex activation was observed in response to pure tone exposure, indicating a music-specific effect. Open-field center crossings were increased concomitant with shifts in music preference, suggesting a potential anxiolytic effect. Thus, music may offer both a unique window into the emotional state of mice and a potentially efficient assay for molecular "brakes" on critical period plasticity common to sensory and higher order brain areas. P reference behaviors are shaped early in life and can last a lifetime. From classic work on chick imprinting (1) to drug vulnerability in adolescents (2, 3), neural circuits in the developing brain are especially impressionable to experience. Biases in response to acoustic signals with ethological or emotional valence over environmental noise are particularly important for social, cultural, and biological fitness. For example, infants will suckle actively to hear their mother's voice over that of another (4) and prefer speech over nonspeech sounds (5) and native over nonnative language (6) as early as 4 d after birth. How and where such enduring preferences are instantiated in the brain remain largely unknown.