Motivation has played an integral role in understanding personality development. Two motivational systems, one associated with seeking reward (approach motivation) and one associated with avoidance of threat (avoidance motivation), have been theorized to represent individual differences in behavioral responses to the environment. However, contextual factors, particularly those with a high degree of novelty, ambiguity, and unpredictability, may simultaneously activate both systems, thereby causing approach-avoidance conflict. The resulting behavior, commonly called inhibition, is characterized by an inability to engage in motivated, goal-directed behavior and is theorized to reflect a core component of anxiety. A form of inhibition observed in childhood, behavioral inhibition (BI), is a relatively stable temperamental profile characterized by negative affect in response to unfamiliar and unpredictable contexts and is a risk factor for anxiety. Our review draws from findings in clinical and cognitive neuroscience to argue that BI reflects an increased sensitivity of both approach and avoidance motivational systems, thereby increasing the likelihood of approach-avoidance conflict within the context of unfamiliar or unpredictable stimuli and environments. Such motivational conflict activates neural systems associated with conflict monitoring, which leads to increases in arousal (e.g., sympathetic nervous system activity) and onlooking behavior, two commonly observed characteristics of childhood BI.