Simultaneous recordings of neuronal activities, pup calls, and maternal behaviors in lactating mice

Maternal behaviors are crucial for the survival and well-being of mammalian offspring. While previous studies have implicated both subcortical and cortical areas, including the auditory cortex (AuCx), in the regulation of maternal behaviors, the real-time dynamics of auditory cortex neuronal activity in response to pup calls during maternal behavior remain unclear. Here, we developed an integrated system that enables simultaneous recording of neural activity, pup calls, and maternal behaviors in freely behaving lactating female mice. Using this system, we observed reliable activity in auditory cortex neurons specifically during the pup-contact phase of retrieval, but not during other maternal behaviors (grooming, crouching, nest-building). Pup calls alone elicited no significant population response in AuCx, as measured by fiber photometry. Furthermore, this activity during pup contact showed a lack of association with the number of pup calls. This study provides an efficient tool for studying neural activity and behavioral dynamics in freely-behaving animals, offering opportunities to investigate the neural mechanisms underlying maternal behaviors. Significance Statement Maternal behaviors are crucial for the survival and well-being of mammalian offspring. In this study, we developed an integrated system that enables simultaneous recording of neural activity, pup calls, and maternal behaviors in freely behaving lactating mice. Using this system, we observed reliable activities in AuCx neurons during pup retrieval. This study provides an efficient tool for studying neural activity and behavioral dynamics in freely behaving animals, offering new opportunities to investigate the neural mechanisms underlying maternal behaviors. Beyond the AuCx, the system could be adapted to investigate the roles of other cortical and deep brain regions in regulating maternal behaviors.