Atrioventricular (AV) delay that results in maximum ventricular filling and physiological mechanisms that govern dependence of filling on timing of atrial systole were studied by combining computer experiments with experiments in the anesthetized dog instrumented to measure phasic mitral flow. Ventricular filling volume is maximized at AV delay of 100 ms in the computer study and 80 ms in the dog study. At any time in diastole atrial contraction accelerates mitral flow, opening the mitral valve widely; atrial relaxation then decelerates mitral flow, moving the valve leaflets toward closure. The time the valve remains closed following atrial systole varies inversely with AV delay. When AV delay is optimal, the mitral valve is moving rapidly toward closure but is not yet closed at onset of ventricular systole. The decline in filling volume as AV delay decreases below its optimum value is primarily the result of premature termination of atrial ejection by ventricular systole. As AV delay increases above its optimal value, filling volume progressively decreases because of premature mitral valve closure that limits effective diastolic filling period. There is no significant retrograde mitral flow at any point in diastole for any AV delay.