Myocardial function is enhanced by endurance exercise training but the cellular mechanisms underlying this improved function remain unclear. The ability of the myocardium to perform external work is a critical aspect of ventricular function, but previous studies of myocardial adaptation to exercise training have been limited to measurements of isometric tension or unloaded shortening velocity, conditions in which work output is zero. We measured force-velocity properties in single permeabilized myocyte preparations in order to determine the effect of exercise training on loaded shortening and power output. Female Sprague-Dawley rats were divided into sedentary control (C) and exercise trained (T) groups. The T rats underwent 11 weeks of progressive treadmill exercise. Myocytes were isolated from T and C hearts, chemically skinned, and attached to a force transducer. Shortening velocity was determined during loaded contractions at 15 °C using a force-clamp technique. Power output was calculated by multiplying force times velocity values. We found that unloaded shortening velocity (V_max) was not significantly different in T vs. C myocytes (T= 1.43 muscle lengths/sec., n = 46 myocytes; C= 1.12 muscle lengths/sec., n= 43 myocytes). Training increased the velocity of loaded shortening and increased peak power output (Peak power = 0.16 P/P_o *ML/sec for T myocytes; Peak power = 0.10 P/P_o *ML/sec for C myocytes). We found no effect of training on myosin heavy chain (MHC) isoform content. These results suggest that training alters power output properties of single cardiac myocytes and this adaptation may improve the work capacity of the myocardium.