Background: Although menopause is known to increase cardiovascular risk and mortality, the effect of menopause on cardiac functions has not been investigated in detail. This study investigates the effect of menopause on cardiac functions by tissue Doppler echocardiography (TDE) and myocardial performance index (MPI). Methods and Results: A total of 72 postmenopausal and 71 age-matched premenopausal women were enrolled in the study. After conventional echocardiographic parameters were measured, TDE recordings were obtained at the septal, lateral, anterior and inferior side of the mitral annulus, and tricuspid lateral annulus. Systolic velocity (Sm), early and late diastolic velocities (Em and Am) and time intervals were measured and MPI was calculated. A sequentially symptom-limited exercise stress test was performed. Although left ventricular (LV) ejection fraction and end-diastolic and end-systolic diameter were similar in both groups, LV septum and posterior wall thickness were higher in postmenopausal women. Mitral early inflow velocity and mitral early inflow velocity: mitral late inflow velocity ratio were significantly lower in postmenopausal women compared to premenopausal women. LV Sm, and LV and right ventricular (RV) Em:Am ratios were lower in postmenopausal women. MPI calculated by TDE was significantly increased in postmenopausal women. In addition, exercise duration and metabolic equivalent values were significantly lower in postmenopausal women than in premenopausal women. Conclusions: Menopause negatively affects MPI and myocardial velocities, both of which provide more quantitative data about myocardial functions. These findings indicate that the hormonal changes in menopause impair LV systolic and diastolic functions and RV diastolic function.