Left ventricular structure has proved to be related with cardiac function; however the relation of myocardial fiber distribution with regional wall motion remains to be elucidated. In this study, both in vivo tagging and ex vivo DTI studies were performed in adult rats. LV circumferential strain, myocardium twist and myocardial fiber architecture were investigated. Results show that myocardial fiber distribution has direct relation with LV myocardium magnitude of circumferential strain and twist angle. Such integrated functional and structural analysis may provide more information for understanding the fundamental cardiac mechanics and assessment of pathological changes.