Infant crying elicits caregiving behaviors, which are indispensable for fulfilling the infant’s needs. However, infant crying can also evoke negative and angry feelings in the caregiver. Therefore, parents need to regulate their own negative emotions to infant crying to sensitively respond to the distressed infant. Thus, the current study aims to examine the neural basis of emotional reactivity and emotional regulation in response to infant crying using functional MRI in primiparous mothers (N = 26). Amygdala activation in response to infant crying was negatively associated with maternal sensitivity and maternal nonhostility during mother–infant interaction. Furthermore, subjective emotional intensity and bilateral activations in the amygdala were decreased using distraction as emotion-regulation strategy. This finding adds neural evidence to the importance of maternal emotion-regulation in response to infant distress. This might be particularly important as some mothers may become overwhelmed by their infant’s distress resulting in intense negative emotions that could potentially impair mother–child interaction and increase child abuse potential.