The purpose of this investigation was to examine longitudinally gestational age and developmental differences in preterm infants' self-regulatory abilities in response to a painful stressor, as well as associations between behavioral and cardiovascular responses. Participants included 49 healthy premature infants. Behavioral and cardiovascular responses to a heel stick blood draw were compared between infants of 28-31 and 32-34 weeks' gestation age at birth. Both gestational age groups displayed behavioral and cardiovascular indications of stress in response to the blood draw. However, both shortly after birth and several weeks later, infants born at younger gestational ages (28-31 weeks) were more physiologically reactive. Evidence that the behavioral stress responses of 28-31 weeks' gestation age group preterm infants do not reflect their physiological responses suggests that evaluation of preterm infants' experiences and risk require assessments of both physiology and behavior. The greater stress vulnerability of the 28-31 weeks' gestation group relative to the 32-34 weeks' gestation group and the implications of this for subsequent development are discussed.