Previous research indicates that maternal cortisol function and maternal brain response to infant are each in turn related to variations in parenting behavior. However, little is known about how maternal cortisol and maternal brain function are associated, thus studying these two mechanisms together may improve our understanding of how maternal cortisol assessed during interactions with own infant is associated with brain response to infant cry. First-time mothers (N = 59) of infants aged 3–4 months old were recruited to participate. Mothers’ cortisol concentration was measured during a naturalistic interaction with their infant and their behavior was coded for two parenting behaviors-- maternal sensitivity and non-intrusiveness. In an fMRI session, mothers listened to their own infant and a control infant crying. Higher cortisol concentration was associated with more intrusive behavior. We found greater cortisol concentration was further associated with decreased activation in the brain to infant cry in the right precentral gyrus, the left culmen extending into the left inferior temporal gyrus and fusiform, two clusters in the superior temporal gyrus, and in the medial frontal gyrus. We also found that lower activation in these regions was associated with more intrusive maternal behavior. These data demonstrate the associations between maternal cortisol concentration and reduced brain activation to infant cry in both motor planning and auditory processing regions in predicting intrusive parenting behavior.