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Individual differences in adolescent exercise behavior are strongly influenced by genetic factors. The affective response to exercise is a potential source of these genetic influences. To test its role in the motivation to exercise, we estimated the heritability of the affective responses during and after exercise and the overlap with the genetic factors influencing regular voluntary exercise behavior.226 twin pairs and 38 siblings completed two submaximal exercise tests on a cycle ergometer and a treadmill and a maximal exercise test on a cycle ergometer. Affective responses were assessed by the Feeling Scale (FS), Borg's Rating of Perceived Exertion (RPE) and the Activation-Deactivation Adjective Checklist (AD ACL).Multivariate structural equation modeling was used to estimate heritability of the affective responses during and after submaximal and maximal exercise and the (genetic) correlation with self-reported regular voluntary exercise behavior over the past year.Genetic factors explained 15% of the individual differences in FS responses during the cycle ergometer test, as well as 29% and 35% of the individual differences in RPE during the cycle ergometer and treadmill tests, respectively. For the AD ACL scales, heritability estimates ranged from 17% to 37% after submaximal exercise and from 12% to 37% after maximal exercise. Without exception, more positive affective responses were associated with higher amounts of regular exercise activity (0.15 < r < 0.21) and this association was accounted for by an overlap in genetic factors influencing affective responding and exercise behavior.We demonstrate low to moderate heritability estimates for the affective response during and after exercise and significant (genetic) associations with regular voluntary exercise behavior. These innate individual differences in the affective responses to exercise should be taken into account in interventions aiming to motivate adolescents to adopt and maintain regular exercise.An important determinant of exercise behavior is the affective response to exercise.Genetic factors explain up until 37% of the variance in this affective response.This affective response shows significant (genetic) associations exercise behavior.