We examined regional brain electrical activity (EEG), heart rate, and subjective responses at rest and during the presentation of videoclips designed to elicit a range of emotions (e.g., sadness, anger, happiness, fear) among a sample of healthy 4‐year‐old children selected for temperamental shyness. We found that shy children exhibited significantly greater relative right central EEG activation at rest and during the presentation of the fear‐eliciting videoclip than nonshy children. Shy females displayed greater relative right mid‐frontal EEG activation during the sad, happy, and fear videoclips than shy males who displayed greater relative left mid‐frontal EEG activation. These results (1) suggest that recent frontal EEG activation/emotion models might be gender‐specific and (2) appear to provide the first empirical evidence for recent theoretical notions linking the origins and maintenance of temperamental shyness in children to difficulty in regulating fear responses. © 2006 Wiley Periodicals, Inc. Dev Psyshobiol 48: 187–196, 2006.